Imidazo-, pyrazolopyrazines and imidazotriazines and their use

ABSTRACT

The invention relates to substituted imidazo-, pyrazolopyrazines and imidazotriazines and processes for their preparation, and their use for the manufacture of medicaments for the treatment and/or prophylaxis of diseases, in particular of hematological disorders, preferably of leukopenias and neutropenias.

The invention relates to substituted imidazo-, pyrazolopyrazines andimidazotriazines and processes for their preparation, and their use forthe manufacture of medicaments for the treatment and/or prophylaxis ofdiseases, in particular of hematological disorders, preferably ofleukopenias and neutropenias.

Glycogen synthase kinase 3 (GSK3) belongs to the families ofserine/threonin kinases. Specific substrates are inter alia cytoskeletalproteins and transcription factors. Two isoforms, GSK3α and GSK3β, havebeen identified to date (Woodgett J R., Trends Biochem. Sci. (1991),16(5), 177-81). Both isoforms are constitutively active in chieflyresting, non-proliferating cells.

GSK3β is of central importance within the Wnt/Wingless signaltransduction pathway. The latter is one of the most important,evolutionarily conserved signalling systems. Wnt signals control veryearly patterning processes during embryogenesis, they induce mesodermformation and many organs, and they control the proliferation anddifferentiation of stem cells (Wodarz A., Nusse R., Annu. Rev. Cell Dev.Biol. (1998), 14, 59-88; Kirstetter et al., Nat. Immunol. (2006), 7(10),1048-56). There is intracellular compartmentalization of the Wntsignalling pathway, thus making it possible to control a wide variety ofprocesses. Within the Wnt cascade, glycogen synthase kinase 3 forms partof a multiprotein complex to which belong inter alia the structuralmolecules axin, the tumor suppressor protein APC and the transcriptioncofactor β-catenin. In this connection, β-catenin is the principalsubstrate of GSK3β. The consequence of this GSK3β-mediatedphosphorylation is the proteasomal degradation of β-catenin. Inhibitionof GSK3 activity leads to an accumulation of β-catenin in the cell withsubsequent translocation into the cell nucleus. There, β-catenin acts asa cofactor in transcription complexes and thus is partly responsible forthe expression of defined target genes.

Radiotherapies or chemotherapies are among the standard approaches tocontrolling cancer. Both types of therapy are nonspecific in relation totheir target cells, i.e. not only tumor cells but also untransformed,proliferating cells are affected. These untransformed, proliferatingcells also include hematopoietic progenitor cells which develop interalia into neutrophilic granulocytes. A significant reduction in thenumber of neutrophiles is referred to as neutropenia. A neutropeniainduced by chemotherapy or radiotherapy results clinically in anincreased susceptibility to infection. If the neutropenia is substantialthere is an increase in the morbidity and, in some circumstances, alsothe mortality of a therapy (O'Brien et al., British Journal of Cancer(2006), 95, 1632-1636).

Inhibition of GSK3 activity leads to an increased rate of proliferationand differentiation of hematopoietic stem cells and can accordingly beutilized for therapeutic intervention in relation to a therapy-inducedneutropenia.

WO99/064401 describes inter alia imidazopyrazines as somatostatinreceptor ligands for the treatment of diabetes. WO2004/026877,US2006/0183746, US2006/0106023 and WO2007/058873 describe the use ofimidazopyrazinylamines for the treatment of cancer. Pyrazolo- andimidazopyrazines for the treatment of cancer are disclosed inWO2006/044687. WO03/000693 claims imidazotriazines as PDE10 inhibitorsfor the treatment of neurodegenerative diseases. WO 2007/145921describes imidazopyrazines as protein kinase inhibitors for thetreatment of cancer.

One object of the present invention is therefore to provide novelcompounds as GSK3β inhibitors for the treatment of hematologicaldisorders, preferably of neutropenia in humans and animals.

The invention provides compounds of the formula

in whicheitherU represents N,V represents CR¹²,W represents CH,A represents CR¹⁵,orU represents CH,V represents CR¹²,W represents N,A represents CR¹⁵,orU represents CR¹⁶,V represents N,W represents CR¹⁷,A represents N,

-   -   where    -   R¹² represents hydrogen, hydroxyl, amino, hydroxycarbonyl,        aminocarbonyl, trifluoromethyl, trifluoromethoxy, cyano,        C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,        C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,        C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino,        C₁-C₄-alkylsulfonylamino, 5- or 6-membered heterocyclylcarbonyl,        —CH₂R¹³ or —CH₂CH₂R¹⁴,        -   where heterocyclylcarbonyl may be substituted by 1 to 3            substituents, where the substituents independently of one            another are selected from the group consisting of halogen,            oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and            C₁-C₄-alkylaminocarbonyl,        -   and        -   where alkoxy, alkylamino, alkylcarbonyl, alkoxycarbonyl,            alkylaminocarbonyl, alkylcarbonylamino and            alkylsulfonylamino may be substituted by a substituent,            where the substituent is selected from the group consisting            of hydroxyl, amino, hydroxycarbonyl, aminocarbonyl,            C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,            C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino, 5- or            6-membered heterocyclyl and phenyl,            -   where phenyl may be substituted by 1 to 3 substituents,                where the substituents independently of one another are                selected from the group consisting of halogen, cyano,                trifluoromethyl, trifluoromethoxy, aminocarbonyl,                C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁₋₄-alkylamino,                C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,                C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino,            -   and            -   where heterocyclyl may be substituted by 1 to 3                substituents, where the consisting of halogen, oxo,                C₁-C₄-alkoxy, C₁-C₄ alkylamino, C₁-C₄-alkylcarbonyl,                C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl        -   and        -   where        -   R¹³ represents hydroxyl, amino, cyano, hydroxycarbonyl,            aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,            C₁-C₄-alkylcarbonylamino, C₃-C₆-cycloalkylamino or 5- or            6-membered heterocyclyl,            -   where alkoxy, alkylamino, alkoxycarbonyl,                alkylaminocarbonyl and alkylcarbonylamino may be                substituted by a substituent, where the substituent is                selected from the group consisting of hydroxyl, amino,                hydroxycarbonyl, aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄                alkylamino, C₁-C₄-alkoxycarbonyl,                C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino,            -   and            -   where heterocyclyl may be substituted by 1 to 3                substituents, where the substituents independently of                one anther are selected from the group consisting of                halogen, oxo, C₁-C₄-alkoxy, C₁-C₄ alkylamino,                C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and                C₁-C₄-alkylaminocarbonyl,        -   and        -   where        -   R¹⁴ represents hydroxyl, amino, cyano, hydroxycarbonyl,            aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,            C₁-C₄-alkylcarbonylamino or 5- or 6-membered heterocyclyl,            -   where alkoxy, alkylamino, alkoxycarbonyl,                alkylaminocarbonyl and alkylcarbonylamino may be                substituted by a substituent, where the substituent is                selected from the group consisting of hydroxyl, amino,                hydroxycarbonyl, aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄                alkylamino, C₁-C₄-alkoxycarbonyl,                C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino,            -   and            -   where heterocyclyl may be substituted by 1 to 3                substituents, where the substituents independently of                one another are selected from the group consisting of                halogen, oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy,                C₁-C₄-alkylamino, C₁-C₄-alkylcarbonyl,                C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl,    -   R¹⁵ represents hydrogen, halogen, cyano, trifluoromethyl,        C₁-C₃-alkyl, methoxy, methylthio or cyclopropyl,    -   R¹⁶ represents hydrogen or methyl,    -   R¹⁷ represents hydrogen or methyl,        R¹ represents a group of the formula

-   -   where    -   * is the point of attachment to the heterocycle,    -   n represents the number 0 or 1,    -   X represents NR¹⁰, S or O,        -   where        -   R¹⁰ represents hydrogen, C₁-C₃-alkyl or cyclopropyl,    -   Y represents NR¹¹ or S,        -   where        -   R¹¹ represents hydrogen, C₁-C₃-alkyl or cyclopropyl,    -   R³ represents pyrid-2-yl, pyrimid-2-yl, 2-aminopyrimid-4-yl,        2-(mono-C₁-C₄-alkylamino)pyrimid-4-yl,        2-(mono-C₃-C₄-cycloalkylamino)pyrimid-4-yl, pyridazin-3        (2H)-on-6-yl, 1,3-oxazol-2-yl, 1,3-oxazol-4-yl,        1,2,4-oxadiazol-3-yl, 1,2,3-oxadiazol-4-yl, 1,3-thiazol-2-yl,        1,3-thiazol-4-yl, 1H-1,2,4-triazol-5-yl,        2,4-dihydro-3H-1,2,4-triazol-3-on-5-yl or 1,2-pyrazol-5-yl,        -   where pyrid-2-yl, pyrimid-2-yl, 1,3-oxazol-2-yl,            1,3-oxazol-4-yl, 1,3-thiazol-2-yl and 1,3-thiazol-4-yl are            substituted by 1 or 2 substituents, where the substituents            independently of one another are selected from the group            consisting of halogen, cyano, nitro, amino, trifluoromethyl,            trifluoromethoxy, aminocarbonyl, trifluoromethylcarbonyl,            C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₃-C₄-cycloalkylamino, C₁-C₄-alkylcarbonyl,            C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl and            C₃-C₆-cycloalkylcarbonyl,            -   where alkyl, alkoxy, alkylamino, alkylcarbonyl,                alkoxycarbonyl, alkylaminocarbonyl and                cycloalkylcarbonyl may be substituted by a substituent,                where the substituent is selected from the group                consisting of halogen, cyano, hydroxyl, amino,                trifluoromethyl and C₃-C₆-cycloalkyl,        -   and        -   where 2-aminopyrimid-4-yl,            2-(mono-C₁-C₄-alkylamino)pyrimid-4-yl,            2-(mono-C₃-C₄-cycloalkylamino)pyrimid-4-yl,            pyridazin-3(2H)-on-6-yl, 1,2,4-oxadiazol-3-yl,            1,2,3-oxadiazol-4-yl, 1H-1,2,4-triazol-5-yl,            2,4-dihydro-3H-1,2,4-triazol-3-on-5-yl and 1,2-pyrazol-5-yl            may be substituted by a substituent, where the substituent            is selected from the group consisting of halogen, cyano,            nitro, amino, trifluoromethyl, trifluoromethoxy,            aminocarbonyl, trifluoromethylcarbonyl, C₁-C₄-alkyl,            C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₃-C₄-cycloalkylamino,            C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl and            C₃-C₆-cycloalkylcarbonyl,    -   R⁴ represents hydrogen, C₁-C₃-alkyl or cyclopropyl,    -   R⁵ represents hydrogen or C₁-C₃-alkyl,    -   R⁶ represents hydrogen, C₁-C₃-alkyl or cyclopropyl,    -   R⁷ represents hydrogen or C₁-C₃-alkyl,    -   R⁸ represents hydrogen, C₁-C₃-alkyl or cyclopropyl,    -   R⁹ represents hydrogen or C₁-C₃-alkyl,        R² represents C₆-C₁₀-aryl or 5- to 10-membered heteroaryl,    -   where aryl and heteroaryl may be substituted by 1 to 3        substituents, where the substituents independently of one        another are selected from the group consisting of hydroxyl,        hydroxymethyl, amino, halogen, cyano, trifluoromethyl,        trifluoromethoxy, aminocarbonyl, C₁-C₄-alkyl, C₁-C₄-alkoxy,        C₁-C₄-alkoxymethyl, C₁-C₄-alkylamino, C₁-C₄-alkylaminomethyl,        C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,        C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino,        C₁-C₄-alkylsulfonyl, C₁-C₄-alkylsulfonylamino,        C₁-C₄-alkylaminosulfonyl, phenyl, benzyloxy, 5- or 6-membered        heterocyclyl, 5- or 6-membered heterocyclylcarbonyl, 5- or        6-membered heterocyclylmethyl and 5- or 6-membered heteroaryl,        -   where phenyl, benzyloxy, heterocyclyl, heterocyclylcarbonyl,            heterocyclylmethyl and heteroaryl may be substituted by 1 to            3 substituents, where the substituents independently of one            another are selected from the group consisting of halogen,            cyano, trifluoromethyl, trifluoromethoxy, aminocarbonyl,            C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,            C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino,    -   or    -   two of the substituents on aryl together with the carbon atoms        to which they are attached form a 1,3-dioxolane or 1,4-dioxane,        and their salts, their solvates and the solvates of their salts.

Compounds according to the invention are the compounds of the formula(I) and the salts, solvates and solvates of the salts thereof, as wellas the compounds encompassed by the formula (I) and mentioned below asexemplary embodiment(s), and the salts, solvates and solvates of thesalts thereof, insofar as the compounds encompassed by formula (I) andmentioned below are not already salts, solvates and solvates of thesalts.

The compounds of the invention may, depending on their structure, existin stereoisomeric forms (enantiomers, diastereomers). The inventiontherefore encompasses the enantiomers or diastereomers and respectivemixtures thereof. The stereoisomerically pure constituents can beisolated in a known manner from such mixtures of enantiomers and/ordiastereomers.

Where the compounds of the invention can occur in tautomeric forms, thepresent invention encompasses all tautomeric forms.

Salts preferred for the purposes of the present invention arephysiologically acceptable salts of the compounds of the invention.However, salts which are themselves unsuitable for pharmaceuticalapplications but can be used for example for isolating or purifying thecompounds of the invention are also encompassed.

Physiologically acceptable salts of the compounds of the inventioninclude acid addition salts of mineral acids, carboxylic acids andsulfonic acids, e.g. salts of hydrochloric acid, hydrobromic acid,sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonicacid, toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonicacid, acetic acid, trifluoroacetic acid, propionic acid, lactic acid,tartaric acid, malic acid, citric acid, fumaric acid, maleic acid andbenzoic acid.

Physiologically acceptable salts of the compounds of the invention alsoinclude salts of conventional bases such as, for example and preferably,alkali metal salts (e.g. sodium and potassium salts), alkaline earthmetal salts (e.g. calcium and magnesium salts) and ammonium saltsderived from ammonia or organic amines having 1 to 16 C atoms, such as,for example and preferably, ethylamine, diethylamine, triethylamine,ethyldiisopropylamine, monoethanolamine, diethanolamine,triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine,dibenzylamine, N-methylmorpholine, arginine, lysine, ethylenediamine,N-methylpiperidine and choline.

Solvates refer for the purposes of the invention to those forms of thecompounds of the invention which form a complex in the solid or liquidstate through coordination with solvent molecules. Hydrates are aspecific form of solvates in which the coordination takes place withwater.

The present invention also encompasses prodrugs of the compoundsaccording to the invention. The term “prodrugs” encompasses compoundswhich themselves may be biologically active or inactive but areconverted during their residence time in the body into compoundsaccording to the invention (for example by metabolism or hydrolysis).

For the purposes of the present invention, the substituents have, unlessspecified otherwise, the following meaning.

Alkyl per se and “alk” and “alkyl” in alkoxy, alkylamino, alkylcarbonyl,alkoxycarbonyl, alkylaminocarbonyl, alkylcarbonylamino, alkylsulfonyl,alkylsulfonylamino and alkylaminosulfonyl stand for a linear or branchedalkyl radical having 1 to 4 carbon atoms, by way of example, andpreferably for methyl, ethyl, n-propyl, isopropyl, n-butyl andtert-butyl.

Alkoxy stands by way of example and preferably for methoxy, ethoxy,n-propoxy, isopropoxy, n-butoxy and tert-butoxy.

Alkylamino stands for an alkylamino radical having one or two alkylsubstituents (chosen independently of one another), by way of exampleand preferably for methylamino, ethylamino, n-propylamino,isopropylamino, tert-butylamino, N,N-dimethylamino, N,N-diethylamino,N-ethyl-N-methylamino, N-methyl-N-n-propylamino,N-isopropyl-N-n-propylamino and N-tert-butyl-N-methylamino.C₁-C₄-alkylamino stands for example for a monoalkylamino radical having1 to 4 carbon atoms or for a dialkylamino radical having 1 to 4 carbonatoms in each alkyl substituent in each case.

Monoalkylamino stands for an alkylamino radical having a linear orbranched alkyl substituent, by way of example and preferably formethylamino, ethylamino, n-propylamino, isopropylamino andtert-butylamino.

Monocycloalkylamino stands for a cycloalkylamino radical having acycloalkyl substituent, where the other substituent at the amino radicalis hydrogen, by way of example and preferably for cyclopropylamino andcyclobutylamino.

Alkylcarbonyl stands by way of example and preferably formethylcarbonyl, ethylcarbonyl, n-propyl-carbonyl, isopropylcarbonyl,n-butylcarbonyl and tert-butylcarbonyl.

Alkoxycarbonyl stands by way of example and preferably formethoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl,n-butoxycarbonyl and tert-butoxycarbonyl.

Alkylaminocarbonyl stands for an alkylaminocarbonyl radical having oneor two alkyl substituents (chosen independently of one another), by wayof example and preferably for methylaminocarbonyl, ethylaminocarbonyl,n-propylaminocarbonyl, isopropylaminocarbonyl, tert-butylaminocarbonyl,N,N-dimethylaminocarbonyl, N,N-diethylaminocarbonyl,N-ethyl-N-methylaminocarbonyl, N-methyl-N-n-propylaminocarbonyl,N-isopropyl-N-n-propylaminocarbonyl andN-tert-butyl-N-methylaminocarbonyl. C₁-C₄-Alkylaminocarbonyl stands forexample for a monoalkylaminocarbonyl radical having 1 to 4 carbon atomsor for a dialkylaminocarbonyl radical having 1 to 4 carbon atoms in eachalkyl substituent in each case.

Alkylcarbonylamino stands by way of example and preferably formethylcarbonylamino, ethyl-carbonylamino, n-propylcarbonylamino,isopropylcarbonylamino, n-butylcarbonylamino andtert-butylcarbonylamino.

Alkylsulfonyl stands by way of example and preferably formethylsulfonyl, ethylsulfonyl, n-propylsulfonyl, isopropylsulfonyl,n-butylsulfonyl and tert-butylsulfonyl.

Alkylaminosulfonyl stands for an alkylaminosulfonyl radical having oneor two alkyl substituents (chosen independently of one another), by wayof example and preferably for methylaminosulfonyl, ethylaminosulfonyl,n-propylaminosulfonyl, isopropylaminosulfonyl, tert-butylaminosulfonyl,N,N-dimethylaminosulfonyl, N,N-diethylaminosulfonyl,N-ethyl-N-methylaminosulfonyl, N-methyl-N-n-propylaminosulfonyl,N-isopropyl-N-n-propylaminosulfonyl andN-tert-butyl-N-methylaminosulfonyl. C₁-C₄-Alkylaminosulfonyl stands forexample for a monoalkylaminosulfonyl radical having 1 to 4 carbon atomsor for a dialkylaminosulfonyl radical having 1 to 4 carbon atoms in eachalkyl substituent in each case.

Alkylsulfonylamino stands by way of example and preferably formethylsulfonylamino, ethyl-sulfonylamino, n-propylsulfonylamino,isopropylsulfonylamino, n-butylsulfonylamino andtert-butylsulfonylamino.

Cycloalkyl stands for a monocyclic cycloalkyl group usually having 3 to6 carbon atoms, and mention may be made by way of example and preferablyof cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl for cycloalkyl.

Cycloalkylamino stands for a cycloalkylamino radical having a cycloalkylsubstituent, the other substituent at the amino radical being hydrogenor an alkyl radical, by way of example and preferably forcyclopropylamino, cyclobutylamino, N-cyclopropyl-N-methylamino andN-cyclobutyl-N-methylamino.

Heterocyclyl stands for a monocyclic, heterocyclic radical having 5 or 6ring atoms and up to 3, preferably up to 2 heteroatoms and/orheterogroups from the series N, O, S, SO, SO₂, where a nitrogen atom mayalso form an N-oxide. The heterocyclyl radicals may be saturated orpartly unsaturated. 5- or 6-membered, monocyclic saturated heterocyclylradicals having up to 2 heteroatoms from the series O, N and S arepreferred, by way of example and preferably for pyrrolidin-2-yl,pyrrolidin-3-yl, pyrrolinyl, tetrahydrofuranyl, tetrahydrothienyl,pyranyl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl,thiopyranyl, morpholin-1-yl, morpholin-2-yl, morpholin-3-yl,piperazin-1-yl, piperazin-2-yl.

Heteroaryl stands for an aromatic, mono- or bicyclic radical usuallyhaving 5 to 10, preferably 5 or 6 ring atoms and up to 5, preferably upto 4 heteroatoms from the series S, O and N, where a nitrogen atom mayalso form an N-oxide, by way of example and preferably for thienyl,furyl, pyrrolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,imidazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, indolyl,indazolyl, benzofuranyl, benzothiophenyl, quinolinyl, isoquinolinyl,benzoxazolyl, benzimidazolyl.

Halogen stands for fluorine, chlorine, bromine and iodine, preferablyfor fluorine and chlorine.

In the formulae of the group which can stand for R¹, the end point ofthe line, besides which a * stands in each case, does not stand for acarbon atom or a CH₂ group but forms part of the bond to the atom towhich R¹ is bonded.

Preference is given to compounds of the formula (I) in which either

U represents N,V represents CR¹²,W represents CH,A represents CR¹⁵,orU represents CH,V represents CR¹²,W represents N,A represents CR¹⁵,orU represents CR¹⁶,V represents N,W represents CR¹⁷,A represents N,

-   -   where    -   R¹² represents hydrogen, hydroxycarbonyl, aminocarbonyl,        C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,        C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,        C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino, 5- or        6-membered heterocyclylcarbonyl, —CH₂R¹³ or —CH₂CH₂R¹⁴,        -   where heterocyclylcarbonyl may be substituted by 1 to 3            substituents, where the substituents independently of one            another are selected from the group consisting of oxo,            C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and            C₁-C₄-alkylaminocarbonyl,        -   and        -   where alkoxy, alkylamino, alkylcarbonyl, alkoxycarbonyl,            alkylaminocarbonyl and alkylcarbonylamino may be substituted            by a substituent, where the substituent is selected from the            group consisting of hydroxyl, amino, hydroxycarbonyl,            aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino and 5- or            6-membered heterocyclyl,            -   where heterocyclyl may be substituted by 1 to 2                substituents, where the substituents independently of                one another are selected from the group consisting of                oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,                C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and                C₁-C₄-alkylaminocarbonyl,        -   and        -   where        -   R¹³ represents hydroxyl, amino, hydroxycarbonyl,            aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,            C₁-C₄-alkylcarbonylamino, C₃-C₆-cycloalkylamino or 5- or            6-membered heterocyclyl,            -   where alkoxy, alkylamino, alkoxycarbonyl,                alkylaminocarbonyl and alkylcarbonylamino may be                substituted by a substituent, where the substituent is                selected from the group consisting of hydroxyl, amino,                hydroxycarbonyl, aminocarbonyl, C₁-C₄-alkoxy,                C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,                C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino,            -   and            -   where heterocyclyl may be substituted by 1 to 2                substituents, where the substituents independently of                one another are selected from the group consisting of                oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,                C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and                C₁-C₄-alkylaminocarbonyl,        -   and        -   where        -   R¹⁴ represents hydroxyl, amino, hydroxycarbonyl,            aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,            C₁-C₄-alkylcarbonylamino or 5- or 6-membered heterocyclyl,            -   where alkoxy, alkylamino, alkoxycarbonyl,                alkylaminocarbonyl and alkylcarbonylamino may be                substituted by a substituent, where the substituent is                selected from the group consisting of hydroxyl, amino,                hydroxycarbonyl, aminocarbonyl, C₁-C₄-alkoxy,                C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,                C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino,            -   and            -   where heterocyclyl may be substituted by 1 to 2                substituents, where the substituents independently of                one another are selected from the group consisting of                oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,                C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and                C₁-C₄-alkylaminocarbonyl,    -   R¹⁵ represents hydrogen, halogen, cyano or trifluoromethyl,    -   R¹⁶ represents hydrogen or methyl,    -   R¹⁷ represents hydrogen or methyl,        R¹ represents a group of the formula

-   -   where    -   * is the point of attachment to the heterocycle,    -   n represents the number 0 or 1,    -   X represents NR¹⁰, S or O,        -   where        -   R¹⁰ represents hydrogen or methyl,    -   Y represents NR¹¹ or S,        -   where        -   R¹¹ represents hydrogen or methyl,    -   R³ represents 2-pyridyl, pyrimid-2-yl, 2-aminopyrimid-4-yl,        1,3-oxazol-2-yl, 1,3-oxazol-4-yl, 1,2,4-oxadiazol-3-yl,        1,2,3-oxadiazol-4-yl, 1,3-thiazol-2-yl or 1,3-thiazol-4-yl,        -   where 2-pyridyl, pyrimid-2-yl, 1,3-oxazol-2-yl,            1,3-oxazol-4-yl, 1,3-thiazol-2-yl and 1,3-thiazol-4-yl are            substituted by 1 or 2 substituents, where the substituents            independently of one another are selected from the group            consisting of halogen, cyano, nitro, amino, trifluoromethyl,            trifluoromethoxy, aminocarbonyl, trifluoromethylcarbonyl            methyl, ethyl, methoxy, ethoxy, C₁-C₄-alkylamino,            methylcarbonyl, ethylcarbonyl, cyclopropylcarbonyl,            methoxycarbonyl and ethoxycarbonyl,        -   and        -   where 2-aminopyrimid-4-yl, 1,2,4-oxadiazol-3-yl and            1,2,3-oxadiazol-4-yl may be substituted by a substituent,            where the substituent is selected from the group consisting            of halogen, cyano, nitro, amino, trifluoromethyl,            trifluoromethoxy, aminocarbonyl, trifluoromethylcarbonyl,            methyl, ethyl, methoxy, ethoxy, C₁-C₄-alkylamino,            methylcarbonyl, ethylcarbonyl, cyclopropylcarbonyl,            methoxycarbonyl and ethoxycarbonyl,    -   R⁴ represents hydrogen or methyl,    -   R⁵ represents hydrogen or methyl,    -   R⁶ represents hydrogen or methyl,    -   R⁷ represents hydrogen or methyl,    -   R⁸ represents hydrogen or methyl,    -   R⁹ represents hydrogen or methyl,

-   R² represents C₆-C₁₀-aryl, thienyl, furyl, pyrrolyl, thiazolyl,    oxazolyl, oxadiazolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidyl,    pyridazinyl, pyrazinyl, indolyl, indazolyl, quinolinyl, benzofuranyl    or benzoxazolyl,    -   where aryl, thienyl, furyl, pyrrolyl, thiazolyl, oxazolyl,        oxadiazolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidyl,        pyridazinyl, pyrazinyl, indolyl, indazolyl, quinolinyl,        benzofuranyl and benzoxazolyl may be substituted by 1 to 3        substituents, where the substituents independently of one        another are selected from the group consisting of hydroxyl,        hydroxymethyl, amino, halogen, cyano, trifluoromethyl,        trifluoromethoxy, aminocarbonyl, C₁-C₄-alkyl, C₁-C₄-alkoxy,        C₁-C₄-alkoxymethyl, C₁-C₄-alkylamino, C₁-C₄-alkylaminomethyl,        C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,        C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino,        C₁-C₄-alkylsulfonyl, C₁-C₄-alkylsulfonylamino,        C₁-C₄-alkylaminosulfonyl, phenyl, benzyloxy, 5- or 6-membered        heterocyclyl, 5- or 6-membered heterocyclylcarbonyl, 5- or        6-membered heterocyclylmethyl and 5- or 6-membered heteroaryl,        -   where phenyl, benzyloxy, heterocyclyl, heterocyclylcarbonyl,            heterocyclylmethyl and heteroaryl may be substituted by 1 to            3 substituents, where the substituents independently of one            another are selected from the group consisting of halogen,            cyano, trifluoromethyl, trifluoromethoxy, aminocarbonyl,            C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,            C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,            C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino,            and their salts, their solvates and the solvates of their            salts.            Preference is also given to compounds of the formula (I) in            which            either            U represents N,            V represents CR¹²,            W represents CH,            A represents CR¹⁵,            or            U represents CH,            V represents CR¹²,            W represents N,            A represents CR¹⁵,            or            U represents CR¹⁶,            V represents N,            W represents CR¹⁷,            A represents N,    -   where    -   R¹² represents hydrogen, hydroxycarbonyl, aminocarbonyl, methyl,        ethyl, C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,        C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino,        pyrrolidinylcarbonyl, piperidinylcarbonyl, piperazinylcarbonyl,        morpholinylcarbonyl or —CH₂R¹³,        -   where pyrrolidinylcarbonyl, piperidinylcarbonyl,            piperazinylcarbonyl and morpholinylcarbonyl may be            substituted by 1 to 2 substituents, where the substituents            independently of one another are selected from the group            consisting of oxo, methyl and ethyl,        -   and        -   where alkylcarbonyl, C₂-C₄-alkoxycarbonyl and            C₂-C₄-alkylaminocarbonyl may be substituted by a            substituent, where the substituent is selected from the            group consisting of hydroxyl, amino, C₁-C₄-alkylamino,            pyrrolidinyl, piperidinyl, piperazinyl and morpholinyl,            -   where pyrrolidinyl, piperidinyl, piperazinyl and                morpholinyl may be substituted by 1 to 2 substituents,                where the substituents independently of one another are                selected from the group consisting of oxo, methyl and                ethyl,        -   and        -   where        -   R¹³ represents hydroxyl, amino, hydroxycarbonyl,            aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, pyrrolidinyl,            piperidinyl, piperazinyl or morpholinyl,            -   where pyrrolidinyl, piperidinyl, piperazinyl and                morpholinyl may be substituted by 1 to 2 substituents,                where the substituents independently of one another are                selected from the group consisting of oxo, methyl and                ethyl,    -   R¹⁵ represents hydrogen,    -   R¹⁶ represents hydrogen or methyl,    -   R¹⁷ represents hydrogen or methyl,        R¹ represents a group of the formula

-   -   where    -   * is the point of attachment to the heterocycle,    -   n represents the number 0,    -   X represents NR¹⁰,        -   where        -   R¹⁰ represents hydrogen,    -   Y represents NR¹¹,        -   where        -   R¹¹ represents hydrogen or methyl,    -   R³ represents 2-pyridyl, pyrimid-2-yl, 2-aminopyrimid-4-yl,        1,3-thiazol-2-yl or 1,3-thiazol-4-yl,        -   where 2-pyridyl, pyrimid-2-yl, 1,3-thiazol-2-yl and            1,3-thiazol-4-yl are substituted by 1 or 2 substituents,            where the substituents independently of one another are            selected from the group consisting of fluorine, chlorine,            cyano, nitro, amino and trifluoromethyl,        -   and        -   where 2-aminopyrimid-4-yl may be substituted by a            substituent, where the substituent is selected from the            group consisting of fluorine, chlorine, cyano, nitro, amino            and trifluoromethyl,    -   R⁴ represents hydrogen,    -   R⁵ represents hydrogen or methyl,    -   R⁶ represents hydrogen,    -   R⁷ represents hydrogen or methyl,    -   R⁸ represents hydrogen,    -   R⁹ represents hydrogen or methyl,        R² represents phenyl, thienyl, pyrazolyl or pyridyl,    -   where phenyl, thienyl, pyrazolyl and pyridyl may be substituted        by 1 to 2 substituents, where the substituents independently of        one another are selected from the group consisting of halogen,        trifluoromethyl, trifluoromethoxy, aminocarbonyl, C₁-C₄-alkyl,        C₁-C₄-alkoxy, C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,        pyrrolidinyl, piperidinyl, morpholinyl and morpholinylcarbonyl,        and their salts, their solvates and the solvates of their salts.

Particular preference is given to compounds of the formula (I) in which

eitherU represents N,V represents CR¹²,W represents CH,A represents CR¹⁵,orU represents CH,V represents CR¹²,W represents N,A represents CR¹⁵,orU represents CR¹⁶,V represents N,W represents CR¹⁷,A represents N,

-   -   where    -   R¹² represents hydrogen, hydroxycarbonyl, methyl, ethyl,        methoxycarbonyl, ethoxycarbonyl, C₁-C₄-alkylaminocarbonyl,        piperidinylcarbonyl or morpholinylcarbonyl,        -   where piperidinylcarbonyl and morpholinylcarbonyl may be            substituted by a substituent, where the substituent is            selected from the group consisting of methyl and ethyl,        -   and        -   where C₂-C₄-alkylaminocarbonyl may be substituted by a            substituent, where the substituent is selected from the            group consisting of C₁-C₄-alkylamino, piperazinyl and            morpholinyl,            -   where piperazinyl and morpholinyl may be substituted by                a substituent, where the substituent is selected from                the group consisting of methyl and ethyl,    -   R¹⁵ represents hydrogen,    -   R¹⁶ represents methyl,    -   R¹⁷ represents methyl,        R¹ represents a group of the formula

-   -   where    -   * is the point of attachment to the heterocycle,    -   n represents the number 0,    -   X represents NR¹⁰,        -   where        -   R¹⁰ represents hydrogen,    -   Y represents NR¹¹,        -   where        -   R¹¹ represents hydrogen or methyl,    -   R³ represents a group of the formula

-   -   -   where        -   # is the point of attachment to Y,        -   L represents cyano, nitro or trifluoromethyl,        -   M represents hydrogen or amino,

    -   R⁴ represents hydrogen,

    -   R⁵ represents hydrogen or methyl,

    -   R⁶ represents hydrogen,

    -   R⁷ represents hydrogen or methyl,

    -   R⁸ represents hydrogen,

    -   R⁹ represents hydrogen,        R² represents phenyl,

    -   where phenyl may be substituted by 1 to 2 substituents, where        the substituents independently of one another are selected from        the group consisting of fluorine, chlorine, trifluoromethyl,        trifluoromethoxy, C₁-C₃-alkyl, methoxy, methoxycarbonyl and        ethoxycarbonyl,        and their salts, their solvates and the solvates of their salts.

Preference is also given to compounds of the formula (I) in which eitherU represents N, V represents CR¹², W represents CH and A represents CHor U represents CH, V represents CR¹², W represents N and A representsCH or U represents CR¹⁶, V represents N, W represents CR¹⁷ and Arepresents N,

where

-   R¹² represents hydrogen, hydroxycarbonyl, methyl, ethyl,    methoxycarbonyl, ethoxycarbonyl, C₁-C₄-alkylaminocarbonyl,    piperidinylcarbonyl or morpholinylcarbonyl,    -   where piperidinylcarbonyl and morpholinylcarbonyl may be        substituted by a substituent, where the substituent is selected        from the group consisting of methyl and ethyl,    -   and    -   where C₂-C₄-alkylaminocarbonyl may be substituted by a        substituent, where the substituent is selected from the group        consisting of C₁-C₄-alkylamino, piperazinyl and morpholinyl,        -   where piperazinyl and morpholinyl may be substituted by a            substituent, where the substituent is selected from the            group consisting of methyl and ethyl,            R¹⁶ represents methyl,            and            R¹⁷ represents methyl.

Preference is also given to compounds of the formula (I) in which Urepresents N, V represents CR¹², W represents CH and A represents CH,

where

-   R¹² represents hydrogen, hydroxycarbonyl, methyl, ethyl,    methoxycarbonyl, ethoxycarbonyl, C₁-C₄-alkylaminocarbonyl,    piperidinylcarbonyl or morpholinylcarbonyl,    -   where piperidinylcarbonyl and morpholinylcarbonyl may be        substituted by a substituent, where the substituent is selected        from the group consisting of methyl and ethyl,    -   and    -   where C₂-C₄-alkylaminocarbonyl may be substituted by a        substituent, where the substituent is selected from the group        consisting of C₁-C₄-alkylamino, piperazinyl and morpholinyl,        -   where piperazinyl and morpholinyl may be substituted by a            substituent, where the substituent is selected from the            group consisting of methyl and ethyl.

Preference is also given to compounds of the formula (I) in which Urepresents CH, V represents CR¹², W represents N and A represents CH,

where

-   R¹² represents hydrogen, hydroxycarbonyl, methyl, ethyl,    methoxycarbonyl, ethoxycarbonyl, C₁-C₄-alkylaminocarbonyl,    piperidinylcarbonyl or morpholinylcarbonyl,    -   where piperidinylcarbonyl and morpholinylcarbonyl may be        substituted by a substituent, where the substituent is selected        from the group consisting of methyl and ethyl,    -   and    -   where C₂-C₄-alkylaminocarbonyl may be substituted by a        substituent, where the substituent is selected from the group        consisting of C₁-C₄-alkylamino, piperazinyl and morpholinyl,        -   where piperazinyl and morpholinyl may be substituted by a            substituent, where the substituent is selected from the            group consisting of methyl and ethyl.

Preference is also given to compounds of the formula (I) in which Urepresents CR¹⁶, V represents N, W represents CR¹⁷ and A represents N,where R¹⁶ and R¹⁷ represents methyl.

Preference is also given to compounds of the formula (I) in which R¹represents —NHCH₂CH₂NH—R³, where R³ represents 5-cyanopyrid-2-yl.

Preference is also given to compounds of the formula (I) in which nrepresents the number 0.

Preference is also given to compounds of the formula (I) in which Xrepresents NR¹⁰, where R¹⁰ represents hydrogen.

Preference is also given to compounds of the formula (I) in which Yrepresents NR¹¹, where R¹¹ represents hydrogen.

Preference is also given to compounds of the formula (I) in which R³represents 5-cyanopyrid-2-yl.

Preference is also given to compounds of the formula (I) in which R⁴,R⁵, R⁶, R⁷, R⁸ and R⁹ represents hydrogen.

The invention furthermore provides a process for preparing the compoundsof the formula (I), or their salts, their solvates or the solvates oftheir salts, where

[A] the compounds of the formula

in whichA, U, V, W and R² have the meaning given above,andX¹ represents halogen, preferably chlorine or fluorine,are reacted with compounds of the formula

R¹—H  (III),

in whichR¹ has the meaning given above,or[B] the compounds of the formula

in whichR¹ has the meaning given above,andA represents CR¹⁵,where R¹⁵ has the meaning given above,U represents N,V represents CR¹²,where R¹² has the meaning given above,W represents CH,X² represents iodine, bromine, chlorine or trifluoromethanesulfonyl,preferably iodine or bromine,are reacted under Suzuki coupling conditions with compounds of theformula

Q—R²  (V),

in whichR² has the meaning given above, andQ represents —B(OH)₂, a boronic acid ester, preferably boronic acidpinacolate, or —BF₃ ⁻K⁺, to give compounds of the formula

in whichR¹ and R² have the meaning given above,andA represents CR¹⁵,where R¹⁵ has the meaning given above,U represents N,V represents CR¹²,where R¹² has the meaning given above,W represents CH.

The compounds of the formula (Ia) are a subset of the compounds of theformula (I).

The reaction according to process [A] is generally carried out in inertsolvents, where appropriate in the presence of a base, where appropriatein a microwave, preferably in a temperature range from 50° C. to 200° C.under atmospheric pressure up to 3 bar.

Examples of bases are alkali metal carbonates, such as, for example,sodium carbonate, potassium carbonate or cesium carbonate, or organicbases, such as trialkylamines, for example triethylamine,N-methylmorpholine, N-methylpiperidine, 4-dimethylaminopyridine ordiisopropylethylamine, or other bases, such as, for example, sodiumhydride or potassium tert-butoxide; preference is given todiisopropylethylamine or sodium hydride.

Examples of inert solvents are halogenated hydrocarbons, such asmethylene chloride or trichloromethane, alcohols, such as methanol,ethanol, n-propanol or isopropanol, or ethers, such as dioxane ortetrahydrofuran, or other solvents, such as, for example, dimethylsulfoxide, dimethylformamide or N-methylpyrrolidone, or mixtures ofthese solvents; preference is given to isopropanol or dimethylsulfoxide.

The reaction according to process [B] is generally carried out in inertsolvents, in the presence of a catalyst, where appropriate in thepresence of an additive, where appropriate in a microwave, preferably ina temperature range from room temperature to 150° C. under atmosphericpressure up to 3 bar.

Examples of catalysts for Suzuki reaction conditions are customarypalladium catalysts; preference is given to catalysts such as, forexample, dichlorobis(triphenylphosphine)palladium,tetrakistriphenylphosphinepalladium(0), palladium(II)acetate/triscyclohexylphosphine,bis-(diphenylphosphaneferrocenyl)palladium(II) chloride,1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene(1,4-naphthtoquinone)palladiumdimer,allyl(chloro)(1,3-dimesityl-1,3-dihydro-2H-imidazol-2-ylidene)palladiumor palladium(II)acetate/dicyclohexyl-(2′,4′,6′-triisopropylbiphenyl-2-yl)phosphine.Another suitable source of palladium istris(dibenzylideneacetone)dipalladium.

Examples of additives are potassium acetate, cesium carbonate, potassiumcarbonate or sodium carbonate, potassium tert-butoxide, cesium fluorideor potassium phosphate carried out; preference is given to additivessuch as, for example, potassium acetate and/or aqueous sodium carbonatesolution.

Examples of inert solvents are ethers, such as dioxane, tetrahydrofuranor 1,2-dimethoxyethane, hydrocarbons, such as benzene, xylene ortoluene, or carboxamides, such as dimethylformamide ordimethylacetamide, alkyl sulfoxides, such as dimethyl sulfoxide, orN-methylpyrrolidone or acetonitrile, or mixtures of the solvents withalcohols, such as methanol or ethanol, and/or water; preference is givento dioxane or acetonitrile or a mixture of one of these solvents withwater.

The compounds of the formulae (II) and (IV) are known, they can besynthesized by known processes from the appropriate starting materialsor they can be prepared analogously to processes described in theexample section (Example 3A to 5A, Example 9A, Example 10A to 12A andExample 13A to 16A) or analogously to J. Org. Chem. (2005), 70 (18),7331-7337 and WO 03/000693.

The compounds of the formula (III) are known, they can be synthesized byknown processes from the appropriate starting materials or they can beprepared analogously to the processes described in the example section(Example 1A to 2A and Example 6A to 8A).

The compounds of the formula (V) are known or they can be synthesized byknown processes from the appropriate starting materials.

The preparation of the starting materials and the compounds of theformula (I) can be illustrated by the synthesis schemes below.

The compounds according to the invention show a valuable range ofpharmacological and pharmacokinetic effects which could not have beenpredicted.

They are therefore suitable of use as medicaments for the treatmentand/or prophylaxis of diseases in humans and animals.

The present invention further relates to the use of the compoundsaccording to the invention for the treatment and/or prophylaxis ofdisorders, preferably hematological disorders, especially of leukopeniasand neutropenia.

The compounds according to the invention are therefore suitable for theprophylaxis and/or treatment of neurodegenerative disorders such as, forexample, Alzheimer's, Parkinson's, schizophrenia, degeneration,dementia, depression, aggression, cerebrovascular ischemia, sleepdisorders, Huntington's chorea, neurotraumatic disorders such as, forexample, stroke; type 2 diabetes mellitus and associated disorders suchas, for example, the metabolic syndrome or obesity, type 1 diabetesmellitus, diabetic nephropathy, diabetic neuropathy, diabeticretinopathy, glomerulonephritis, hypercalcemia, hyperglycemia,hyperlipidemia, glucose-galactose malabsorption, general endocrinedysfunctions such as, for example, pancreatitis; hematological disorderssuch as, for example, acquired and congenital neutropenia,medicament-induced neutropenia, parasite-induced neutropenia,chemotherapy-induced neutropenia, granulocytopenia, acquired andcongenital leukopenia, acquired and congenital anemia, hemolytic anemia,sickle cell anemia, acquired and congenital thrombocytopenia, leukocytedysfunctions, impairments of blood coagulation, ex vivo expansion ofembryonic and adult stem cells, ex vivo differentiation of embryonic andadult stem cells, bone marrow, graft-versus-host reaction; cancer suchas, for example, glaucoma, breast carcinoma, colon tumor,gastrointestinal tumors, Hodgkin's lymphoma, non-Hodgkin's lymphoma,Kaposi sarcoma, liver tumor, pancreatic tumor, skin tumor, bone marrowtumor, leukemias such as, for example, acute lymphatic leukemia, acutemyeloid leukemia, chronic myeloid leukemia, chronic lymphatic leukemia,prostate tumors, lung cancer, renal tumors; asthma, progressive, notcompletely reversible obstruction of the respiratory tract, pneumonia,pulmonary dysfunction; inflammatory disorders such as, for example,autoimmune diseases such as multiple sclerosis, rheumatoid arthritis,infections by gram-negative and gram-positive bacteria, viralinfections, fungal infections such as, for example, by Candida albicans,HIV infections and HIV-associated infections, hepatitis of types A, Band C, parasitic infections; hair loss; reduced sperm mobility; woundhealing; osteoporosis, bone marrow disorders, bone and joint disorders;cardiovascular disorders such as, for example, cardiac defects, heartfailure, cardiac fibrosis, cardiac arrhythmias, myocardial infarction,medicament- or substance-induced cardiotoxicity, atherosclerosis, highblood pressure.

The compounds according to the invention are particularly suitable forthe prophylaxis and/or treatment of neurodegenerative disorders, suchas, for example, Alzheimer's disease and schizophrenia, of type IIdiabetes mellitus and associated disorders, of cancer, of leukopeniasand/or of neutropenias.

The compounds according to the invention can additionally be employedalso for efficient ex vivo expansion of adult hematopoietic stem cellsfrom the bone marrow, from peripheral blood or from umbilical cordblood.

These cells expanded in this way can then be used to curtail thecytopenias induced by myeloablative therapies or within the framework oftherapeutic transplantation methods or for hematological systemicdisorders such as, for example, leukemias, or with cells which have beengenetically manipulated after expansion for gene therapies.

The present invention further relates to the use of the compoundsaccording to the invention for the treatment and/or prophylaxis ofdisorders, especially of the aforementioned disorders.

The present invention further relates to the use of the compoundsaccording to the invention for the manufacture of a medicament for thetreatment and/or prophylaxis of disorders, especially of theaforementioned disorders.

The present invention further relates to a method for the treatmentand/or prophylaxis of disorders, in particular of the aforementioneddisorders, by use of a therapeutically effective amount of a compoundaccording to the invention.

The present invention further relates to medicaments comprising acompound according to the invention and one or more further activeingredients.

The present invention further relates to a method for the ex vivoexpansion of adult hematopoietic stem cells from bone marrow, fromperipheral blood or from umbilical cord blood, which is characterized inthat an effective amount of the compound according to the invention isadded.

The compounds of the invention can act systemically and/or locally. Forthis purpose, they can be administered in a suitable way such as, forexample, by the oral, parenteral, pulmonary, nasal, sublingual, lingual,buccal, rectal, dermal, transdermal, conjunctival or otic route or asimplant or stent.

The compounds of the invention can be administered in administrationforms suitable for these administration routes.

Suitable for oral administration are administration forms which functionaccording to the prior art and deliver the compounds of the inventionrapidly and/or in modified fashion, and which contain the compounds ofthe invention in crystalline and/or amorphized and/or dissolved form,such as, for example, tablets (uncoated or coated tablets, for examplehaving enteric coatings or coatings which are insoluble or dissolve witha delay and control the release of the compound of the invention),tablets which disintegrate rapidly in the mouth, or films/wafers,films/lyophilizates, capsules (for example hard or soft gelatincapsules), sugar-coated tablets, granules, pellets, powders, emulsions,suspensions, aerosols or solutions.

Parenteral administration can take place with avoidance of an absorptionstep (e.g. intravenous, intraarterial, intracardiac, intraspinal orintralumbar) or with inclusion of an absorption (e.g. intramuscular,subcutaneous, intracutaneous, percutaneous or intraperitoneal).Administration forms suitable for parenteral administration are, interalia, preparations for injection and infusion in the form of solutions,suspensions, emulsions, lyophilizates or sterile powders.

Oral administration is preferred.

Suitable for the other administration routes are, for example,pharmaceutical forms for inhalation (inter alia powder inhalers,nebulizers), nasal drops, solutions, sprays; tablets for lingual,sublingual or buccal administration, films/wafers or capsules,suppositories, preparations for the ears or eyes, vaginal capsules,aqueous suspensions (lotions, shaking mixtures), lipophilic suspensions,ointments, creams, transdermal therapeutic systems (such as, forexample, patches), milk, pastes, foams, dusting powders, implants orstents.

The compounds of the invention can be converted into the statedadministration forms. This can take place in a manner known per se bymixing with inert, non-toxic, pharmaceutically suitable excipients.These excipients include, inter alia, carriers (for examplemicrocrystalline cellulose, lactose, mannitol), solvents (e.g. liquidpolyethylene glycols), emulsifiers and dispersants or wetting agents(for example sodium dodecyl sulfate, polyoxysorbitan oleate), binders(for example polyvinylpyrrolidone), synthetic and natural polymers (forexample albumin), stabilizers (e.g. antioxidants such as, for example,ascorbic acid), colorants (e.g. inorganic pigments such as, for example,iron oxides) and masking flavours and/or odours.

The present invention further relates to medicaments which comprise atleast one compound of the invention, preferably together with one ormore inert, non-toxic, pharmaceutically suitable excipients, and to theuse thereof for the aforementioned purposes.

It has generally proved advantageous on parenteral administration toadminister amounts of about 5 to 500 mg every 24 hours to achieveeffective results. The amount on oral administration is about 5 to 500mg every 24 hours.

It may nevertheless be necessary where appropriate to deviate from thestated amounts, in particular as a function of the body weight, route ofadministration, individual response to the active ingredient, nature ofthe preparation and time or interval over which administration takesplace.

The percentage data in the following tests and examples are, unlessindicated otherwise, percentages by weight; parts are parts by weight.Solvent ratios, dilution ratios and concentration data of liquid/liquidsolutions are in each case based on volume. The statement “w/v” means“weight/volume”. Thus, for example, “10% w/v” means: 100 ml of solutionor suspension comprise 10 g of substance.

A) EXAMPLES Abbreviations

abs. absoluteBoc tert-butoxycarbonylCDCl₃ deuterochloroformconc. concentratedd day

DIEA N,N-diisopropylethylamine DMAP 4-N,N-dimethylaminopyridine

DMF dimethylformamideDMSO dimethyl sulfoxideEDC N′-(3-dimethylaminopropyl)-N-ethylcarbodiimide x HCleq. equivalentESI electrospray ionization (in MS)h hourHOBt 1-hydroxy-1H-benzotriazole x H₂OHPLC high pressure, high performance liquid chromatographyLC-MS coupled liquid chromatography-mass spectrometrymin. minutesMS mass spectrometryMW molecular weight [g/mol]NMR nuclear magnetic resonance spectroscopyOAc acetateOEt ethoxyp.a. per analysisPyBOP 1-benzotriazolyloxytripyrrolidinophosphonium hexafluorophosphateRf retention index (in TLC)sat. saturatedRP-HPLC reverse phase HPLCRT room temperatureR_(t) retention time (in HPLC)TBTU (benzotriazol-1-yloxy)bisdimethylaminomethylium fluoroborateTFA trifluoroacetic acidTHF tetrahydrofuran

LC-MS Methods:

Method 1: Instrument: Micromass Quattro LCZ with HPLC Agilent series1100; column: Phenomenex Synergi 2.5μ MAX-RP 100 A mercury 20 mm×4 mm;eluent A: 1 l of water+0.5 ml of 50% formic acid, eluent B: 1 l ofacetonitrile+0.5 ml of 50% formic acid; gradient: 0.0 min 90% A→0.1 min90% A→3.0 min 5% A→4.0 min 5% A→4.1 min 90% A; flow rate: 2 ml/min;oven: 50° C.; UV detection: 208-400 nm.

Method 2: MS instrument type: Micromass ZQ; HPLC instrument type: WatersAlliance 2795; column: Merck Chromolith SpeedROD RP-18e 100 mm×4.6 mm;eluent A: water+500 μl of 50% formic acid/1; eluent B: acetonitrile+500μl of 50% formic acid/1; gradient: 0.0 min 10% B→7.0 min 95% B→9.0 min95% B; oven: 35° C.; flow rate: 0.0 min 1.0 ml/min→7.0 min 2.0ml/min→9.0 min 2.0 ml/min; UV detection: 210 nm

Method 3: MS instrument type: Micromass ZQ; HPLC instrument type: HP1100 Series; UV DAD; column: Phenomenex Gemini 3μ 30 mm×3.00 mm; eluentA: 1 l of water+0.5 ml of 50% formic acid, eluent B: 1 l ofacetonitrile+0.5 ml of 50% formic acid; gradient: 0.0 min 90% A→2.5 min30% A→3.0 min 5% A→4.5 min 5% A; flow rate: 0.0 min 1 ml/min, 2.5min/3.0 min/4.5 min. 2 ml/min; oven: 50° C.; UV detection: 210 nm.

Method 4: Instrument: Micromass Platform LCZ with HPLC Agilent series1100; column: Thermo Hypersil GOLD 3μ 20 mm×4 mm; eluent A: 1 l ofwater+0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile+0.5 ml of50% formic acid; gradient: 0.0 min 100% A→0.2 min 100% A→2.9 min 30%A→3.1 min 10% A→5.5 min 10% A; oven: 50° C.; flow rate: 0.8 ml/min; UVdetection: 210 nm.

Method 5: MS instrument type: Waters ZQ; HPLC instrument type: WatersAlliance 2795; column: Phenomenex Onyx Monolithic C18, 100 mm×3 mm;eluent A: 1 l of water+0.5 ml of 50% formic acid, eluent B: 1 l ofacetonitrile+0.5 ml of 50% formic acid; gradient 0.0 min 90% A→2 min 65%A→4.5 min 5% A→6 min 5% A; flow rate: 2 ml/min; oven: 40° C.; UVdetection: 210

Method 6: MS instrument type: Micromass ZQ; HPLC instrument type: WatersAlliance 2795; column: Phenomenex Synergi 2.5μ MAX-RP 100 A mercury 20mm×4 mm; eluent A: 1 l of water+0.5 ml of 50% formic acid, eluent B: 1 lof acetonitrile+0.5 ml of 50% formic acid; gradient: 0.0 min 90% A→0.1min 90% A→3.0 min 5% A→4.0 min 5% A→4.01 min 90% A; flow rate: 2 ml/min;oven: 50° C.; UV detection: 210 nm.

Method 7: Instrument: Micromass Quattro LCZ with HPLC Agilent series1100; column: Phenomenex Onyx Monolithic C18, 100 mm×3 mm; eluent A: 1 lof water+0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile+0.5 mlof 50% formic acid; gradient: 0.0 min 90% A→2 min 65% A→4.5 min 5% A→6min 5% A; flow rate: 2 ml/min; oven: 40° C.; UV detection: 208-400 nm.

Method 8: Instrument: Micromass QuattroPremier with Waters UPLC Acquity;column: Thermo Hypersil GOLD 1.9μ 50 mm×1 mm; eluent A: 1 l of water+0.5ml of 50% formic acid, eluent B: 1 l of acetonitrile+0.5 ml of 50%formic acid; gradient: 0.0 min 100% A→0.1 min 100% A→1.5 min 10% A→2.2min 10% A; oven: 50° C.; flow rate: 0.33 ml/min; UV detection: 210 nm.

Method 9: Instrument: Micromass Quattro Micro MS with HPLC Agilentseries 1100; column: Thermo Hypersil GOLD 3μ 20 mm×4 mm; eluent A: 1 lof water+0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile+0.5 mlof 50% formic acid; gradient: 0.0 min 100% A→3.0 min 10% A→4.0 min 10%A→4.01 min 100% A→5.00 min 100% A; flow rate: 0.0 min/3.0 min/4.0min/4.01 min 2.5 ml/min, 5.00 min 2 ml/min; oven: 50° C.; UV detection:210 nm.

Methode 10: MS instrument: Waters ZQ 2000; HPLC instrument: Agilent1100, 2-column setup, autosampler: HTC PAL; column: YMC-ODS-AQ, 50mm×4.6 mm, 3.0 μm; eluent A: water+0.1% formic acid, eluent B:acetonitrile+0.1% formic acid; gradient: 0.0 min 100% A→0.2 min 95%A→1.8 min 25% A→1.9 min 10% A→2.0 min 5% A→3.2 min 5% A→3.21 min 100%A→3.35 min 100% A; oven: 40° C.; flow rate: 3.0 ml/min; UV detection:210 nm.

Methode 11: MS instrument: Micromass TOF (LCT); HPLC instrument: Waters2690, autosampler: Waters 2700; column: YMC-ODS-AQ, 50 mm×4.6 mm, 3.0μm; eluent A: water+0.1% formic acid, eluent B: acetonitrile+0.1% formicacid; gradient: 0.0 min 100% A→0.2 min 95% A→1.8 min 25% A→1.9 min 10%A→2.0 min 5% A→3.2 min 5% A→3.21 min 100% A→3.35 min 100% A; oven: 40°C.; flow rate: 3.0 ml/min; UV detection: 210 nm.

Starting Materials Example 1A tert-Butyl{2-[(5-cyanopyridin-2-yl)-amino]ethyl}carbamate

5.5 g (39.7 mmol) of 6-chloronicotinonitrile were dissolved in 70 ml ofDMSO, and 10.2 g (63.5 mmol) of N-Boc-ethylenediamine and 11 g (79.4mmol) of potassium carbonate were added. The mixture was stirred at 90°C. for 12 h. The residue was taken up in a mixture of water and ethylacetate. The organic phase was washed with saturated aqueous sodiumchloride solution, dried over magnesium sulfate and concentrated on arotary evaporator. The residue was chromatographed on silica gel 60(mobile phase: cyclohexane/ethyl acetate 10:1 to 2:1). This gave 7.9 g(77% of theory) of the product as a solid.

LCMS (method 6): R_(t)=1.46 min. (m/z=263 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.37 (d, 1H), 7.66 (d, 1H) 7.6 (s, 1H),6.87 (t, 1H), 6.53 (d, 1H), 3.32 (q, 2H), 3.09 (q, 2H), 1.37 (s, 9H).

Example 2A 6-[(2-Aminoethyl)amino]nicotinonitrile dihydrochloride

7.9 g (30 mmol) of tert-butyl{2-[(5-cyanopyridin-2-yl)amino]ethyl}carbamate (Example 1A) weredissolved in 100 ml of 4N hydrogen chloride in dioxane and the mixturewas stirred for 30 min. The reaction mixture was concentrated to half ofits original volume and the same amount of diethyl ether was added. Thereaction mixture was stirred for 20 min and the product was filtered offand washed with diethyl ether. This gave 7 g (94% of theory) of theproduct as a solid.

LCMS (method 4): R_(t)=0.51 min. (m/z=162 (M+H)±)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.44 (s, 1H), 7.76 (d, 1H), 6.67 (d, 1H),3.58 (t, 2H), 2.98 (q, 2H).

Example 3A Ethyl1-[2-(2,4-dichlorophenyl)-2-oxoethyl]-1H-imidazole-2-carboxylate

0.5 g (3.6 mmol) of imidazole-2-carboxylate ethyl were dissolved in 35ml of acetone, and 0.96 g (3.6 mmol) of 2-bromo-2,4-dichloroacetophenoneand 0.49 g (3.6 mmol) of potassium carbonate were added. The mixture wasstirred at RT for 12 h. The reaction mixture was concentrated and takenup in water and dichloromethane. The organic phase was washed withsaturated aqueous sodium chloride solution, dried over magnesium sulfateand concentrated on a rotary evaporator. The residue was triturated withdiethyl ether and the solid was filtered off with suction. This gave 0.9g (77% of theory) of the product as a solid.

LCMS (method 1): R_(t)=1.19 min. (m/z=327 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=7.96 (d, 1H), 7.84 (d, 1H), 7.68 (dd, 1H),7.5 (s, 1H), 7.16 (s, 1H), 5.87 (s, 2H), 4.22 (q, 2H), 3.32 (s, 2H),1.23 (t, 3H).

Example 4A 6-(2,4-Dichlorophenyl)imidazo[1,2-a]pyrazin-8(7H)-one

920 mg (2.8 mmol) of ethyl1-[2-(2,4-dichlorophenyl)-2-oxoethyl]-1H-imidazol-2-carboxylate (Example3A) were dissolved in 45 ml of glacial acetic acid, and 2.17 g (28 mmol)of ammonium acetate were added. The mixture was stirred under reflux for12 h. The reaction mixture was added to ice-water and neutralized andwith sodium carbonate. The precipitate was filtered off and dried underhigh vacuum. This gave 650 mg (82% of theory) of the product as a solid.

LCMS (method 3): R_(t)=1.79 min. (m/z=281 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=11.5 (s, 1H), 7.83 (s, 1H) 7.81 (d, 1H),7.63 (s, 1H), 7.61 (s, 1H), 7.58 (d, 1H), 7.52 (s, 1H).

Example 5A 8-Chloro-6-(2,4-dichlorophenyl)imidazo[1,2-a]pyrazine

650 mg (2.3 mmol) of6-(2,4-dichlorophenyl)imidazo[1,2-a]pyrazin-8(7H)-one (Example 4A) weredissolved in 7 ml of phosphoryl chloride and stirred under reflux for 12h. The reaction mixture was poured into 100 ml of saturated sodiumbicarbonate solution, and solid sodium bicarbonate was added until a pHof 7 had been reached. Ethyl acetate was added, and the mixture wasextracted. The organic phase was washed with saturated aqueous sodiumchloride solution, dried over magnesium sulfate and concentrated on arotary evaporator. Methanol was added to the residue, and theprecipitated solid was filtered off and washed with methanol and diethylether and dried under high vacuum. This gave 400 mg (58% of theory) ofthe product as a solid.

LCMS (method 3): R_(t)=2.43 min. (m/z=299 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=9.01 (s, 1H), 8.34 (s, 1H) 7.93 (s, 1H),7.81 (d, 1H), 7.72 (d, 1H), 7.6 (dd, 1H).

Example 6A 4-Amino-2-(methylsulfonyl)-1,3-thiazole-5-carbonitrile

2.7 g (15.8 mmol) of4-amino-2-(methylsulfanyl)-1,3-thiazole-5-carbonitrile were dissolved in500 ml of dichloromethane, and 12 g (34.7 mmol) of 50% strength3-chloroperbenzoic acid were added. The mixture was stirred at RT for 30min. 6 ml of DMSO were added, and the reaction mixture was washed withsaturated sodium bicarbonate solution and water. The organic phase wasdried over magnesium sulfate and concentrated on a rotary evaporator.This gave 2.2 g (46% of theory) of the product as a solid.

LCMS (method 4): R_(t)=1.19 min. (m/z=204 (M+H)⁺).

Example 7A tert-Butyl{2-[(4-amino-5-cyano-1,3-thiazol-2-yl)amino]ethyl}carbamate

3.25 g (16 mmol) of4-amino-2-(methylsulfonyl)-1,3-thiazole-5-carbonitrile (Example 6A) weredissolved in 50 ml of DMSO, and 3.8 g (24 mmol) of N-Boc-ethylenediamineand 2.8 ml (16 mmol) of N,N-diisopropylethylamine were added. Themixture was stirred at 120° C. for 12 h. The reaction mixture was takenup in a mixture of ethyl acetate and water. The organic phase was washedwith saturated aqueous sodium chloride solution, dried over magnesiumsulfate and concentrated on a rotary evaporator. The crude product wastriturated with acetonitrile and the precipitated crystals were filteredoff with suction. This gave 2.5 g (55% of theory) of the product as asolid.

LCMS (method 4): R_(t)=1.81 min. (m/z=284 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=8.35 (s, 1H), 6.9 (t, 1H), 6.68 (s, 2H),3.22 (q, 2H), 3.07 (q, 2H), 1.36 (s, 9H).

Example 8A 4-Amino-2-[(2-aminoethyl)amino]-1,3-thiazol-5-carbonitriledihydrochloride

2.18 g (7.7 mmol) of tert-butyl{2-[(4-amino-5-cyano-1,3-thiazol-2-yl)amino]ethyl}carbamate (Example 7A)in 100 ml of 4N hydrogen chloride in dioxane were stirred at RT for 30min. The reaction mixture was concentrated. This gave 2 g (100% oftheory) of the product as a solid.

LCMS (method 6): R_(t)=0.19 min. (m/z=183 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=3.46 (d, 2H), 3.0 (d, 2H).

Example 9A4-Amino-2-({2-[(6-bromoimidazo[1,2-a]pyrazin-8-yl)amino]ethyl}amino)-1,3-thiazole-5-carbonitrile

300 mg (1.08 mmol) of 6,8-dibromoimidazo[1,2-a]pyrazine were dissolvedin 10 ml of DMSO, and 0.6 ml (4.3 mmol) of triethylamine and 360 mg (1.3mmol) of 4-amino-2-[(2-aminoethyl)amino]-1,3-thiazole-5-carbonitriledihydrochloride (Example 8A) were added. The mixture was heated at 140°C. in a microwave for 1 h. Purification by preparative HPLC gave 376 mg(83% of theory) of the product as a solid.

LCMS (method 8): R_(t)=0.88 min. (m/z=381 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=8.46 (t, 1H), 8.12 (t, 1H), 8.05 (s, 1H),7.85 (s, 1H), 7.53 (s, 1H), 3.6 (d, 2H), 3.48 (d, 2H).

Example 10A Diethyl1-[2-(2,4-dichlorophenyl)-2-oxoethyl]-1H-pyrazole-3,5-dicarboxylate

5 g (23.6 mmol) of diethyl 1H-pyrazole-3,5-dicarboxylate were dissolvedin 100 ml of acetone, and 6.3 g (23.6 mmol) of2-bromo-2,4-dichloroacetophenone and 3.6 g (26 mmol) of potassiumcarbonate were added. The mixture was stirred at RT for 12 h. Thereaction mixture was concentrated and the residue was taken up in waterand dichloromethane. The organic phase was washed with saturated aqueoussodium chloride solution, dried over magnesium sulfate and concentratedon a rotary evaporator. The residue was triturated with diethyl ether,and the solid was filtered off with suction. This gave 9.48 g (94% oftheory) of the product as a solid.

LCMS (method 6): R_(t)=2.34 min. (m/z=399 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.02 (d, 1H), 7.84 (s, 1H), 7.68 (dd, 1H),7.35 (s, 1H), 6.13 (s, 2H), 4.32 (q, 2H), 4.26 (q, 2H), 1.31 (t, 3H),1.25 (t, 3H).

Example 11A Ethyl6-(2,4-dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carboxylate

9.5 g (23.7 mmol) of diethyl1-[2-(2,4-dichlorophenyl)-2-oxoethyl]-1H-pyrazole-3,5-dicarboxylate(Example 10A) were dissolved in 300 ml of glacial acetic acid, and 18.3g (237 mmol) of ammonium acetate were added. The mixture was stirred atreflux for 12 h. The reaction mixture was poured into ice-water andneutralized with sodium carbonate. The precipitate was filtered off anddried under high vacuum. This gave 6.86 g (82% of theory) of the productas a solid.

LCMS (method 3): R_(t)=2.31 min (m/z=354 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=11.85 (s, 1H), 7.98 (s, 1H) 7.83 (d, 1H),7.64 (d, 1H), 7.59 (dd, 1H), 7.44 (s, 1H), 4.34 (q, 2H), 1.33 (t, 3H).

Example 12A Ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate

6.86 g (19.5 mmol) of ethyl6-(2,4-dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carboxylate(Example 11A) were dissolved in 50 ml of phosphoryl chloride and stirredat reflux for 12 h. The reaction mixture was poured into 1.25 l ofsaturated sodium bicarbonate solution, and solid sodium bicarbonate wasadded until a pH of 7 had been reached. The solid was filtered off anddissolved in dichloromethane and chromatographed on silica gel (mobilephase cyclohexane/ethyl acetate 10:1). This gave 6.34 g (85% of theory)of the product as a solid.

LCMS (method 3): R_(t)=3.15 min. (m/z=371 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=9.3 (s, 1H), 7.84 (d, 1H) 7.73 (d, 1H),7.62 (dd, 1H), 7.56 (s, 1H), 4.4 (q, 2H), 1.36 (t, 3H).

Example 13A Ethyl 3-acetamido-2-oxobutanoate

52 g (396 mmol) of acetylalanine were dissolved in 800 ml of THF, and 45mg (0.4 mmol) of 4-dimethylaminopyridine and 96 ml (1188 mmol) ofpyridine were added. The mixture was heated to reflux, and 88 ml (792mmol) of ethyl oxalyl chloride were added dropwise over a period of 45min. The mixture was stirred at reflux for a further 3 h. The reactionmixture was added to ice-water, ethyl acetate was added and the mixturewas extracted. The organic phase was washed with saturated aqueoussodium chloride solution, dried over sodium sulfate and concentrated ona rotary evaporator. The crude mixture was reacted further without anyfurther purification.

¹H-NMR (400 MHz, CDCl₃): δ=10.86 (s, 1H), 4.42 (q, 1H) 4.23 (q, 2H),2.16 (s, 3H), 1.42 (t, 3H), 1.29 (t, 3H).

Example 14AN-[1-(5-Oxo-3-phenyl-4,5-dihydro-1,2,4-triazin-6-yl)ethyl]acetamide

48 g (306 mmol) of benzenecarboximidamide hydrochloride were dissolvedin 1.2 l ethanol, and 16.4 ml (337 mmol) of hydrazine hydrate wereadded. The mixture was stirred at 45° C. for 3 h. 86 g (459 mmol) ofethyl 3-acetamido-2-oxobutanoate (Example 13A), dissolved in 600 ml ofethanol, were added dropwise. The mixture was stirred at 80° C. foranother 6 h and at RT for another 12 h. The reaction mixture waschromatographed on silica gel (mobile phase dichloromethane/methanol20:1). This gave 23.2 g (26% of theory) of the product as a solid.

¹H-NMR (400 MHz, CDCl₃): δ=8.21 (d, 2H), 7.56 (m, 3H) 5.36 (q, 1H), 2.05(s, 3H), 1.55 (d, 3H).

Example 15A 5,7-Dimethyl-2-phenylimidazo[5,1-f][1,2,4]triazin-4(3H)-one

8.34 g (32.3 mmol) ofN-[1-(5-oxo-3-phenyl-4,5-dihydro-1,2,4-triazin-6-yl)ethyl]acetamide](Example 14A) were dissolved in 330 ml of 1,2-dichloroethane and 4.5 ml(48.5 mmol) of phosphoryl chloride were added. The mixture was stirredunder reflux for 24 h. After cooling, the precipitate was filtered offand washed with water and diethyl ether. The product was subsequentlydried under high vacuum. This gave 4.6 g (59% of theory) of the productas a solid.

¹H-NMR (400 MHz, DMSO-d₆): δ=12.45 (s, 1H), 8.08 (d, 2H), 7.6 (m, 3H),2.69 (s, 3H), 2.59 (s, 3H).

Example 16A 4-Chloro-5,7-dimethyl-2-phenylimidazo[5,1-f][1,2,4]triazine

700 mg (2.9 mmol) of5,7-dimethyl-2-phenylimidazo[5,1-f][1,2,4]triazin-4(3H)-one (Example15A) were dissolved in 8 ml of phosphoryl chloride and stirred at refluxfor 12 h. The reaction mixture was poured into 170 ml of saturatedsodium bicarbonate solution, and solid sodium bicarbonate was addeduntil a pH of 7 had been reached. Ethyl acetate was added, and themixture was extracted. The organic phase was washed with saturatedaqueous sodium chloride solution, dried over magnesium sulfate andconcentrated on a rotary evaporator. This gave 657 mg (80% of theory) ofthe product as a solid.

LCMS (method 6): R_(t)=2.17 min. (m/z=259 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=8.26 (dd, 2H), 7.58 (m, 3H), 2.68 (s, 3H),2.66 (s, 3H).

Example 17A6-(2,4-Dichlorophenyl)-2-(hydroxymethyl)pyrazolo[1,5-a]pyrazin-4(5H)-one

5 g (12.55 mmol) of ethyl6-(2,4-dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carboxylate(Example 11A) were dissolved in 700 ml of THF, and 952 mg (25.1 mmol) oflithium aluminum hydride were added at RT. The mixture was stirred at RTfor 2 h, after complete conversion methanol was added first, and the pHwas then adjusted to 6 using dilute hydrochloric acid. The mixture wasextracted repeatedly with ethyl acetate. Removal of the solvent gave3.12 g (80% of theory) of the product as a solid.

LCMS (method 3): R_(t)=1.66 min. (m/z=310 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=11.55 (s, 1H), 7.81 (d, 1H), 7.77 (s, 1H),7.62 (d, 1H), 7.66 (dd, 1H), 6.94 (s, 1H), 5.32 (t, 1H), 4.58 (d, 2H).

Example 18A6-(2,4-Dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carbaldehyde

1.15 g (3.37 mmol) of6-(2,4-dichlorophenyl)-2-(hydroxymethyl)pyrazolo[1,5-a]pyrazin-4(5H)-one(Example 17A) were suspended in 100 ml of dichloromethane, and 1 drop ofwater was added. 100 ml of 1,2-dimethoxyethane and 9.6 ml of DMF werethen added, and the mixture was cooled to 0° C. 7.16 g (16.87 mmol) ofDess-Martin periodinane were added, ice-bath cooling was removed and themixture was stirred at RT for 15 h. Ethyl acetate and then 200 ml of a10% strength sodium thiosulfate solution were added to the mixture, andthe organic phase was washed with saturated aqueous sodium bicarbonatesolution. Drying of the organic phase with sodium sulfate and removal ofthe solvent under reduced pressure gave 660 mg (57% of theory) of theproduct as a solid which was reacted without any further purification.

LCMS (method 8): R_(t)=1.04 min. (m/z=308 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=11.89 (s, 1H), 10.07 (s, 1H), 8.02 (s, 1H),7.84 (d, 1H), 7.66 (d, 1H), 7.60 (dd, 1H), 7.52 (s, 1H).

Example 19A6-(2,4-Dichlorophenyl)-2-[(4-methylpiperazin-1-yl)methyl]pyrazolo[1,5-a]pyrazin-4(5H)-one

145 mg (0.32 mmol) of6-(2,4-dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carb-aldehyde(Example 18A) were dissolved in 6 ml of methanol, and 64 mg (0.64 mmol)of 1-methylpiperazine, 4 Å molecular sieve and 58 mg (0.96 mmol) ofacetic acid were added. Finally, 40.2 mg (0.64 mmol) of sodiumcyanoborohydride were added and the mixture was stirred at RT for 15 h.The crude mixture was acidified with 2N hydrochloric acid and theresulting precipitate was filtered off with suction. This gave 120 mg(96% of theory) of the product as a solid which was reacted furtherwithout any further purification.

LCMS (method 6): R_(t)=0.76 min. (m/z=392 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=11.63 (s, 1H), 7.82 (d, 1H), 7.81 (s, 1H),7.61 (d, 1H), 7.57 (dd, 1H), 7.0 (s, 1H), 3.7-4.0 (m, 4H), 3.38 (m, 2H),3.04 (m, 4H), 2.78 (s, 3H).

Example 20A4-Chloro-6-(2,4-dichlorophenyl)-2-[(4-methylpiperazin-1-yl)methyl]pyrazolo[1,5-a]pyrazine

Analogously to the preparation of Example 16A, 160 mg (0.34 mmol) of6-(2,4-dichlorophenyl)-2-[(4-methylpiperazin-1-yl)methyl]pyrazolo[1,5-a]pyrazin-4(5H)-onegave, by reaction with phosphoryl chloride, 108 mg (70% of theory) ofthe product as a solid.

LCMS (method 6): R_(t)=0.76 min. (m/z=392 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=9.12 (s, 1H), 7.81 (d, 1H), 7.71 (d, 1H),7.60 (dd, 1H), 6.99 (s, 1H), 3.74 (s, 2H), 2.4-2.55 (m, 4H), 2.25-2.4(m, 4H), 2.15 (s, 3H).

Example 21A6-(2,4-Dichlorophenyl)-2-(morpholin-4-ylmethyl)pyrazolo[1,5-a]pyrazin-4(5H)-onetrifluoroacetate

760 mg (2.17 mmol) of6-(2,4-dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazin-2-carb-aldehyde(Example 18A) were dissolved in 46 ml of methanol, and 378 mg (4.34mmol) of morpholine, 4 Å molecular sieve and 0.373 ml (6.51 mmol) ofacetic acid were added. Finally, 272.8 mg (4.34 mmol) of sodiumcyanoborohydride were added and the mixture was stirred at RT for 15 h.The crude mixture was acidified with 2N hydrochloric acid and theresulting precipitate was filtered off with suction. Purification of theprecipitate by preparative HPLC (mobile phase: acetonitrile/watergradient with 0.1% trifluoroacetic acid) gave 693 mg (65% of theory) ofthe product as a solid.

LCMS (method 8): R_(t)=0.75 min. (m/z=379 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=11.79 (s, 1H), 10.31 (s, br, 1H), 7.91 (s,1H), 7.84 (d, 1H), 7.64 (d, 1H), 7.59 (dd, 1H), 7.22 (s, 1H), 4.54 (s,2H), 3.96 (m, 2H), 3.64 (m, 2H), 3.39 (m, 2H), 3.18 (m, 2H).

Example 22A4-Chloro-6-(2,4-dichlorophenyl)-2-(morpholin-4-ylmethyl)pyrazolo[1,5-a]pyrazinehydrochloride

Analogously to the preparation of Example 16A, 690 mg (1.18 mmol) of6-(2,4-dichlorophenyl)-2-(morpholin-4-ylmethyl)pyrazolo[1,5-a]pyrazin-4(5H)-onetrifluoroacetate gave, by reaction with phosphoryl chloride, 395 mg (85%of theory) of the product as a solid.

LCMS (method 6): R_(t)=1.32 min. (m/z=397 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=11.50 (s, 1H), 9.23 (s, 1H), 7.84 (d, 1H),7.72 (d, 1H), 7.62 (dd, 1H), 7.44 (s, br, 1H), 4.65 (s, br, 2H), 3.9-4.0(m, 2H), 3.7-3.85 (m, 2H), 3.3-3.45 (m, 2H), 3.1-3.25 (m, 2H).

Example 23A6-({2-[(6-Bromoimidazo[1,2-a]pyrazin-8-yl)amino]ethyl}amino)pyridine-3-carbonitrile

8 g (28.9 mmol) of 6,8-dibromoimidazo[1,2-a]pyrazine were dissolved in80 ml of DMSO, and 16.1 ml (115.6 mmol) of triethylamine and 5.154 g(31.78 mmol) of 6-[(2-aminoethyl)-amino]pyridine-3-carbonitrile (Example2A) were added. The mixture was heated at 140° C. in a microwave for 1.5h. The solution was poured into water and the precipitate was filteredoff. Washing with water and drying under high vacuum gave 10.3 g (94% oftheory) of the product as a solid.

LCMS (method 8): R_(t)=0.97 min. (m/z=358 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=8.39 (s, 1H), 8.08 (s, br, 1H), 8.03 (s,1H), 7.83 (s, 1H), 7.73 (m, 2H), 7.51 (s, 1H), 6.56 (s, br, 1H),3.5-3.65 (m, 4H).

Example 24A tert-Butyl (6-chloropyridin-2-yl)carbamate

Under argon, 150 ml of THF were added to 23.4 g (181.8 mmol) of2-chloro-5-aminopyridine and the mixture was cooled to 0° C. 73.3 g (400mmol) of bis(trimethylsilyl)sodium amide and 43.65 g (200 mmol) ofdi-tert-butyl dicarbonate, dissolved in 150 ml of THF, were addeddropwise. After 15 min, the cooling bath was removed and the mixture wasstirred for a further 15 min at RT. The THF was removed using a rotaryevaporator, ethyl acetate and 0.5 N hydrochloric acid were added and themixture was extracted. The organic phase was separated off, dried overmagnesium sulfate and concentrated on a rotary evaporator. The reactionmixture was chromatographed on silica gel (mobile phasedichloromethane/methanol 100%→100:3). This gave 36.54 g (88% of theory)of the product as a solid.

LCMS (method 3): R_(t)=2.41 min. (m/z=175 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=10.11 (s, 1H), 7.78 (d, 2H), 7.1 (t, 1H),1.47 (s, 9H).

Example 25A tert-Butyl (6-chloro-3-formylpyridin-2-yl)carbamate

The reaction apparatus was dried by heating, and the reaction wascarried out under argon with stirring. 15 g (65.6 mmol) of tert-butyl(6-chloropyridin-2-yl)carbamate (Example 24A) and 19 g (164 mmol) of1,2-bis(dimethylamino)ethane were initially charged in 270 ml of THF andcooled to −78° C. 102.5 ml (164 mmol) of butyllithium (1.6N) were addeddropwise. After the dropwise addition had ended, the reaction was slowlywarmed to −10° C. and kept at −10° C. for 2 h. The mixture was then oncemore cooled to −78° C., and 10 ml (131 mmol) of DMF were added. Thereaction was slowly warmed to RT, the reaction mixture was added to 1 lof ethyl acetate and 350 ml of 1N hydrochloric acid and stirred for 15min and the organic phase was separated off. The organic phase waswashed with water and saturated sodium bicarbonate solution, dried overmagnesium sulfate and concentrated on a rotary evaporator. Diethyl etherwas added to the residue, and the solid was filtered off with suctionand dried. This gave 12.3 g (73% of theory) of the product as a solid.

LCMS (method 3): R_(t)=2.19 min. (m/z=255 (M+H)⁻).

¹H-NMR (400 MHz, DMSO-d₆): δ=10.37 (s, 1H), 9.83 (s, 1H), 8.2 (d, 1H),7.42 (d, 1H), 1.46 (s, 9H).

Example 26A tert-Butyl{6-chloro-3-[(hydroxyimino)methyl]pyridin-2-yl}carbamate

15.45 g (60.2 mmol) of tert-butyl(6-chloro-3-formylpyridin-2-yl)carbamate (Example 25A) were initiallycharged in 750 ml of ethanol, a solution of 225 ml of water and 9.38 g(120.4 mmol) of sodium acetate was added and the mixture was stirred for5 min. A solution of 225 ml of water and 8.36 g (114.4 mmol) ofhydroxylamine hydrochloride was added and the mixture was stirred at RTfor 4 h. The reaction mixture was concentrated on a rotary evaporator at20° C. The residue was taken up in ethyl acetate, washed twice withsaturated sodium bicarbonate solution and once with saturated sodiumchloride solution. The organic phase was separated off, dried overmagnesium sulfate and concentrated on a rotary evaporator at 20° C. Thisgave 15.5 g (80% of theory) of the product as a solid.

LCMS (method 3): R_(t)=2.08 min. (m/z=270 (M+H)⁻).

¹H-NMR (400 MHz, DMSO-d₆): δ=11.71 (s, 1H), 9.91 (s, 1H), 8.14 (s, 1H),8.02 (d, 1H), 7.3 (d, 1H), 1.49 (s, 9H).

Example 27A 2-Amino-6-chloropyridine-3-carbaldehyde oxime hydrochloride

15.5 g (57 mmol) of tert-butyl{6-chloro-3-[(hydroxyimino)methyl]pyridin-2-yl}carbamate (Example 26A)were dissolved in 285 ml of 4N hydrogen chloride in dioxane and themixture was stirred for 30 min. The reaction mixture was concentrated tohalf of its original volume and the same amount of diethyl ether wasadded. The reaction mixture was stirred for 20 min and the product wasfiltered off and washed with diethyl ether. This gave 11 g (94% oftheory) of the product as a solid.

LCMS (method 6): R_(t)=1.09 min. (m/z=172 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.27 (s, 1H), 7.61 (d, 1H), 6.65 (d, 1H).

Example 28A 2-Amino-6-chloropyridine-3-carbonitrile

11.15 g (53.6 mmol) of 2-amino-6-chloropyridine-3-carbaldehyde oximehydrochloride (Example 27A) were initially charged in dioxane, 13 ml(161 mmol) of pyridine were added and the mixture was cooled to 0° C.8.3 ml (58.95 mmol) of trifluoroacetic anhydride were added, and thereaction was warmed to RT and then stirred at 60° C. for 2 h. Thereaction mixture was taken up in a mixture of ethyl acetate and sodiumbicarbonate solution. The organic phase was washed with saturatedaqueous sodium chloride solution, dried over magnesium sulfate andconcentrated on a rotary evaporator. The residue was suspended indichloromethane:diethyl ether 3:1, and the solid was filtered off withsuction and dried. This gave 5.56 g (66% of theory) of the product as asolid.

LCMS (method 6): R_(t)=1.0 min. (m/z=154 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=7.91 (d, 1H), 7.38 (s, 2H), 6.69 (d, 1H).

Example 29A tert-Butyl{2-[(6-amino-5-cyanopyridin-2-yl)-amino]ethyl}carbamate

2 g (13 mmol) of 2-amino-6-chloropyridine-3-carbonitrile (Example 28A)were initially charged in 15 ml of DMSO, and 2.71 g (16.93 mmol) ofN-Boc-ethyleneamine and 3.4 ml (19.54 mmol) of N,N-diisopropylethylaminewere added. The reaction mixture was irradiated in a microwave reactorat 115° C. for 1.5 h. The reaction mixture was taken up in a mixture ofethyl acetate and water. The organic phase was washed with saturatedaqueous sodium chloride solution, dried over magnesium sulfate andconcentrated on a rotary evaporator. This gave 23.38 g (88% of theory)of the product as a solid.

LCMS (method 3): R_(t)=1.7 min. (m/z=278 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=7.3 (s, 1H), 7.0 (br, s, 1H), 6.83 (s, 1H),6.25 (s, 2H), 5.78 (d, 1H), 3.25 (q, 2H), 3.06 (q, 2H), 1.36 (s, 9H).

Example 30A 2-Amino-6-[(2-aminoethyl)amino]pyridine-3-carbonitriledihydrochloride

6.76 g (24.38 mmol) of tert-butyl{2-[(6-amino-5-cyanopyridin-2-yl)amino]ethyl}carbamate (Example 29A)were dissolved in 122 ml of a 4N solution of hydrogen chloride indioxane and the mixture was stirred for 30 min. The reaction mixture wasconcentrated to half of its original volume and the same amount ofdiethyl ether was added. The reaction mixture was stirred for 20 min andthe product was filtered off and washed with diethyl ether. This gave5.43 g (89% of theory) of the product as a solid.

LCMS (method 6): R_(t)=0.92 min. (m/z=177 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.1 (s, 2H), 7.5 (d, 1H), 5.96 (d, 1H),3.53 (q, 2H), 3.0 (q, 2H).

Example 31A 4-(Trifluoroacetyl)morpholine

15 g (172 mmol) of morpholine were initially charged in 750 ml ofdichloromethane, and 29 ml (206 mmol) of trifluoroacetic anhydride and119 ml (688 mmol) of N,N-diisopropylethylamine were added at 0° C. Thereaction mixture was warmed to RT and stirred at RT for another 3 h. Thereaction mixture was concentrated and the residue was taken up in ethylacetate and washed successively with aqueous sodium bicarbonatesolution, 1N hydrochloric acid and once more with aqueous sodiumbicarbonate solution. The organic phase was dried over magnesium sulfateand concentrated on a rotary evaporator. This gave 28 g (88% of theory)of the product as an oil.

LCMS (method 9): R_(t)=1.22 min. (m/z=184 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=3.65 (m, 2H), 3.56 (m, 2H).

Example 32A tert-Butyl[6-chloro-3-(trifluoroacetyl)pyridin-2-yl]carbamate

8 g (35 mmol) of tert-butyl (6-chloropyridin-2-yl)carbamate (Example24A) were initially charged in 100 ml of THF and cooled to −50° C. 55 ml(87 mmol) of butyllithium (1.6N) were added dropwise. After the dropwiseaddition had ended, the reaction was slowly warmed to −10° C. andstirred at 0° C. for 2 h. The mixture was then cooled again to −40° C.,and 12.8 g (70 mmol) of 4-(trifluoroacetyl)morpholine (Example 21A),dissolved in 4 ml of THF, were added. The reaction solution was stirredat −40° C. for 1 h, and, at −40° C., poured into 1 l of ethyl acetateand 350 ml of ammonium chloride solution and extracted. The organicphase was separated off, dried over magnesium sulfate and concentratedon a rotary evaporator. The reaction mixture was chromatographed onsilica gel (mobile phase cyclohexane/ethyl acetate 10:1). This gave 9 g(79% of theory) of the product as an oil.

¹H-NMR (400 MHz, DMSO-d₆): δ=10.96 (s, 1H), 7.99 (d, 1H), 7.4 (d, 1H),1.43 (s, 9H).

Example 33A tert-Butyl[6-({2-[(tert-butoxycarbonyl)amino]ethyl}amino)-3-(trifluoroacetyl)pyridin-2-yl]carbamate

5 g (15.4 mmol) of tert-butyl[6-chloro-3-(trifluoroacetyl)pyridin-2-yl]carbamate (Example 32A) wereinitially charged in 37.5 ml of DMSO, and 3.2 g (20 mmol) ofN-Boc-ethylenediamine and 4 ml (23 mmol) of N,N-diisopropylethylaminewere added. The reaction mixture was irradiated in a microwave reactorat 90° C. for 0.5 h. The reaction mixture was taken up in a mixture ofethyl acetate and water. The organic phase was washed with saturatedaqueous sodium chloride solution, dried over magnesium sulfate andconcentrated on a rotary evaporator. The reaction mixture waschromatographed on silica gel (mobile phase cyclohexane/ethyl acetate5:1→1:1). This gave 2.5 g (34% of theory) of the product as a solid.

LCMS (method 6): R_(t)=2.44 min. (m/z=449 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=10.75 (s, 1H), 8.44 (s, 1H), 7.70 (d, 1H),6.77 (s, 1H), 6.28 (d, 1H), 3.48 (br, s, 2H), 3.17 (br, s, 2H), 1.46 (s,9H), 1.30 (s, 9H).

Example 34A1-{2-Amino-6-[(2-aminoethyl)amino]pyridin-3-yl}-2,2,2-trifluoroethanonehydrochloride

2.5 g (5.57 mmol) of tert-butyl[6-({2-[(tert-butoxycarbonyl)amino]ethyl}amino)-3-(trifluoroacetyl)-pyridin-2-yl]carbamate(Example 33A) were dissolved in 15 ml of a 4N solution of hydrogenchloride in dioxane, and the mixture was stirred for 20 h. The reactionmixture was concentrated to half of its original volume and the sameamount of diethyl ether was added. The reaction mixture was stirred for20 min and the product was filtered off and washed with diethyl ether.This gave 1.4 g (89% of theory) of the product as a solid.

LCMS (method 6): R_(t)=0.73 min. (m/z=249 (M+H)⁺).

Example 35A tert-Butyl3-[(5-cyanopyridin-2-yl)-amino]piperidine-1-carboxylate

1.0 g (4.99 mmol) of tert-butyl 3-aminopiperidine-1-carboxylate and1.383 g (9.99 mmol) of 6-chloropyridine-3-carbonitrile and 1.29 g (9.99mmol) of diisopropylethylamine were suspended in 40 ml of DMSO andheated in a microwave at 140° C. for 45 min. Most of the DMSO wasremoved from the mixture by kugelrohr distillation, water was added andthe resulting precipitate was filtered off. Drying under high vacuumgave 2.24 g (46% of theory) of the product.

LCMS (method 3): R_(t)=2.23 min. (m/z=303 (M+H)⁺).

Example 36A 6-(Piperidin-3-ylamino)pyridine-3-carbonitrile hydrochloride

2.24 g (3.4 mmol) of tert-butyl3-[(5-cyanopyridin-2-yl)amino]piperidine-1-carboxylate (Example 35A)were dissolved in 4.3 ml of a solution of hydrochloric acid in dioxane(4 M), and the mixture was stirred at RT for 3 h. After the reaction hadgone to completion, the solvent was removed completely. This gave 1.74 g(90% of theory) of the product as a solid.

LCMS (method 8): R_(t)=0.27 min. (m/z=203 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=9.13 (m, 1H), 9.0 (m, 1H), 8.44 (d, 1H),7.89 (m, 1H), 7.74 (dd, 1H), 6.63 (d, 1H), 5.58 (s, br), 4.19 (s, br,1H), 3.57 (s, 1H), 3.34 (d, 1H), 3.14 (d, 1H), 2.88 (m, 1H), 2.7-2.81(m, 1H), 1.82-2.0 (m, 2H), 1.63-1.79 (m, 1H), 1.48-1.59 (m, 1H).

Example 37A tert-Butyl3-[(6-amino-5-cyanopyridin-2-yl)amino]piperidine-1-carboxylate

2.15 g (10.7 mmol) of tert-butyl 3-aminopiperidine-1-carboxylate, 1.50 g(9.77 mmol) of 2-amino-6-chloropyridine-3-carbonitrile (Example 28A) and1.89 g (14.7 mmol) of diisopropylethylamine were suspended in 6 ml ofDMSO and heated in a microwave reactor at 130° C. for 8 h. The reactionmixture was diluted with ethyl acetate (100 ml) and water (40 ml), andthe organic phase was separated off and washed with saturated aqueoussodium chloride solution (50 ml), dried over magnesium sulfate andconcentrated. The residue was chromatographed on silica gel (mobilephase: cyclohexane/ethyl acetate 4:1 to 1:1). This gave 2.04 g (60% oftheory) of the product as a solid.

LCMS (method 6): R_(t)=1.69 min. (m/z=318 (M+H)⁺)

Example 38A 2-Amino-6-(piperidin-3-ylamino)pyridine-3-carbonitrilehydrochloride

2.00 g (6.3 mmol) of tert-butyl3-[(6-amino-5-cyanopyridin-2-yl)amino]piperidine-1-carboxylate (Example37A) were dissolved in 40 ml of a solution of hydrochloric acid indioxane (4 M), and the mixture was stirred at RT for 2 h. After thereaction had gone to completion the solvent was concentrated to half ofits original volume and 20 ml of diethyl ether were added. Theprecipitate was filtered off and dried. This gave 1.80 g (100% oftheory) of the product as a solid.

LCMS (method 8): R_(t)=0.25 min. (m/z=218 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=9.38 (br m, 1H), 8.97 (br m, 1H), 8.25 (brm, 1H), 7.53 (m, 1H), 7.40 (br s, 2H), 6.01 (d, 1H), 4.16 (br m, 1H),3.34 (br m, 1H), 3.10 (m, 1H), 2.89 (m, 2H), 2.00-1.84 (m, 2H), 1.73 (m,1H), 1.55 (m, 1H).

Example 39A tert-Butyl3-({6-[(tert-butoxycarbonyl)amino]-5-(trifluoroacetyl)pyridin-2-yl}amino)piperidine-1-carboxylate

561 mg (2.8 mmol) of tert-butyl 3-aminopiperidine-1-carboxylate, 700 mg(2.16 mmol) of tert-butyl[6-chloro-3-(trifluoroacetyl)pyridin-2-yl]carbamate (Example 32A) and0.56 ml (3.23 mmol) of diisopropylethylamine were suspended in 14 ml ofDMSO and heated in a microwave reactor at 90° C. for 45 min. Thereaction mixture was diluted with ethyl acetate (100 ml) and washed withsaturated aqueous ammonium chloride solution (three times 40 ml) andthen saturated aqueous sodium bicarbonate solution (40 ml). The organicphase was dried over magnesium sulfate and concentrated. The residue waschromatographed on silica gel (mobile phase: cyclohexane/ethyl acetate5:1 to 1:1). This gave 670 mg (63% of theory) of the product.

LCMS (method 6): R_(t)=2.70 min. (m/z=489 (M+H)⁺)

Example 40A1-[2-Amino-6-(piperidin-3-ylamino)pyridin-3-yl]-2,2,2-trifluoroethanonehydrochloride

670 mg (1.37 mmol) of tert-butyl3-({6-[(tert-butoxycarbonyl)amino]-5-(trifluoroacetyl)pyridin-2-yl}amino)piperidine-1-carboxylate (Example 39A) were dissolved in 25 ml ofa solution of hydrochloric acid in dioxane (4 M), and the mixture wasstirred at RT for 20 h. After the reaction had gone to completion, thereaction mixture was diluted with diethyl ether (100 ml) and theprecipitate was filtered off and washed with diethyl ether (100 ml) anddried. This gave 286 mg (64% of theory) of the product as a solid.

LCMS (method 6): R_(t)=0.81 min. (m/z=289 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=9.26 (br s, 1H), 9.07 (br s, 1H), 8.8.34(br s, 1H), 7.59 (d, 1H), 6.22 (br, 2H), 6.03 (d, 1H), 4.25 (br m, 1H),3.36 (m, 1H), 3.13 (m, 1H), 2.93 (m, 2H), 2.00-1.85 (m, 2H), 1.73 (m,1H), 1.56 (m, 1H).

Example 41A tert-Butyl3-[(6-amino-5-nitropyridin-2-yl)amino]piperidine-1-carboxylate

500 mg (2.11 mmol) of tert-butyl 3-aminopiperidine-1-carboxylate, 772 mg(4.22 mmol) of 2-amino-6-chloro-3-nitropyridine and 1.05 ml (6.34 mmol)of diisopropylethylamine were suspended in 18 ml of DMSO and heated in amicrowave reactor at 120° C. for 45 min. The reaction mixture waspurified by preparative reverse-phase HPLC. This gave 600 mg (81% oftheory) of the product as a solid.

LCMS (method 6): R_(t)=1.77 min. (m/z=338 (M+H)⁺)

Example 42A 3-Nitro-N⁶-(piperidin-3-yl)pyridine-2,6-diaminehydrochloride

610 mg (1.62 mmol) of tert-butyl3-[(6-amino-5-nitropyridin-2-yl)amino]piperidine-1-carboxylate (Example41A) were dissolved in 40 ml of a solution of hydrochloric acid indioxane (4 M), and the mixture was stirred at RT for 30 min. After thereaction had gone to completion, the solvent was removed completely.This gave 662 mg of the crude product.

LCMS (method 4): R_(t)=0.86 min. (m/z=238 (M+H)⁺)

Example 43A Methyl 4-amino-2-(methylsulfonyl)-1,3-thiazole-5-carboxylate

5.12 g (8.32 mmol) of Oxone were dissolved in 170 ml of water and cooledto 5° C. A solution of 1 g (4.90 mmol) of methyl4-amino-2-(methylsulfanyl)-1,3-thiazole-5-carboxylate in 18 ml ofmethanol was then added dropwise, and the solution was stirred at RT for3 h. Most of the methanol was removed, and the remainder was extractedthree times with dichloromethane. The combined organic phases were driedwith sodium sulfate. Following removal of the solvent and drying of theresidue under high vacuum, the solid obtained (824 mg (43% of theory))was used without further purification.

LCMS (method 3): R_(t)=1.52 min. (m/z=237 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=7.36 (s, br, 2H), 3.79 (s, 3H), 3.45 (s,3H).

Example 44A tert-Butyl3-{[4-amino-5-(methoxycarbonyl)-1,3-thiazol-2-yl]amino}piperidine-1-carboxylate

Analogously to the preparation of Example 49A, 335 mg (1.42 mmol) oftert-butyl 3-aminopiperidine-1-carboxylate hydrochloride and 3190 mg(2.84 mmol) of methyl4-amino-2-(methylsulfonyl)-1,3-thiazole-5-carboxylate gave 158 mg (29%of theory) of the product as a solid.

LCMS (method 8): R_(t)=1.06 min. (m/z=357 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=8.32 (d, 1H), 6.79 (s, br, 2H), 3.60 (s,3H), 3.55 (m, 2H), 1.89 (m, 1H), 1.71 (m, 1H), 1.5 (m, 1H), 1.35 (s,11H).

Example 45A Methyl4-amino-2-(piperidin-3-ylamino)-1,3-thiazole-5-carboxylatedihydrochloride

Analogously to the preparation of Example 38A, 150 mg (0.39 mmol) oftert-butyl3-{[4-amino-5-(methoxycarbonyl)-1,3-thiazol-2-yl]amino}piperidine-1-carboxylateand 20 ml of hydrochloric acid in dioxane (4M) gave 130 mg (99% oftheory) of the product as a solid.

LCMS (method 8): R_(t)=0.26 min. (m/z=257 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=9.04 (s, br, 2H), 8.51 (d, 1H), 3.9 (m,2H), 3.6 (s, 3H), 3.34 (d, 1H), 3.11 (d, 1H), 2.75-2.94 (m, 2H),1.93-2.04 (m, 1H), 1.8-1.91 (m, 1H), 1.60-1.76 (m, 1H), 1.43-1.57 (m,1H).

Example 46A tert-Butyl3-[(4-amino-5-cyano-1,3-thiazol-2-yl)amino]piperidine-1-carboxylate

643 mg (3.17 mmol) of4-amino-2-(methylsulfonyl)-1,3-thiazole-5-carbonitrile (Example 6A) weredissolved in 16 ml of DMSO, and 500 mg (2.11 mmol) of tert-butyl3-aminopiperidine-1-carboxylate and 3.49 ml (21.12 mmol) ofN,N-diisopropylethylamine were added. The mixture was heated in amicrowave at 120° C. for 45 min. The reaction mixture was taken up in amixture of ethyl acetate and water. The organic phase was washed withsaturated aqueous sodium chloride solution, dried over magnesium sulfateand concentrated on a rotary evaporator. The crude product was usedwithout further purification.

LCMS (method 8): R_(t)=1.0 min. (m/z=357 (M+H)⁺).

Example 47A 4-Amino-2-(piperidin-3-ylamino)-1,3-thiazole-5-carbonitriledihydrochloride

Analogously to the preparation of Example 38A, 240 mg (0.7 mmol) oftert-butyl3-[(4-amino-5-cyano-1,3-thiazol-2-yl)-amino]piperidine-1-carboxylate(Example 46A) and 25 ml of hydrochloric acid in dioxane (4M) gave 265 mg(57% of theory) of the product as a solid.

LCMS (method 9): R_(t)=0.74 min. (m/z=224 (M+H)⁺).

Example 48A 1-[4-Amino-2-(methylsulfonyl)-1,3-thiazol-5-yl]ethanone

2.775 g (4.52 mmol) of Oxone were dissolved in 9 ml of water, and 500 mg(2.66 mmol) of 1-[4-amino-2-(methylsulfanyl)-1,3-thiazol-5-yl]ethanone,dissolved in 9 ml of methanol, were added dropwise at 5° C. The mixturewas stirred at RT for 3 h, the methanol was reduced on a rotaryevaporator and the residue was extracted twice with dichloromethane.Removal of the solvent gave 395 mg (52% of theory) of the product as asolid.

LCMS (method 3): R_(t)=1.17 min. (m/z=221 (M+H)⁺).

Example 49A tert-Butyl3-[(5-acetyl-4-amino-1,3-thiazol-2-yl)amino]piperidine-1-carboxylate

390 mg (1.77 mmol) of1-[4-amino-2-(methylsulfonyl)-1,3-thiazol-5-yl]ethanone (Example 48A)were dissolved in 5 ml of DMSO, and 209.6 mg (0.89 mmol) of tert-butyl3-aminopiperidine-1-carboxylate and 0.585 ml (3.54 mmol) ofN,N-diisopropylethylamine were added. The mixture was heated in amicrowave at 120° C. for 45 min. This gave, after purification bypreparative HPLC, 162 mg (52% of theory) of the product as a solid.

LCMS (method 8): R_(t)=1.87 min. (m/z=341 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=8.56 (d, 1H), 7.70 (s, br, 2H), 3.81 (m,4H), 3.56 (m, 2H), 2.02 (s, 3H), 1.91 (m, 1H), 1.71 (m, 1H), 1.51 (m,1H), 1.36 (s, 9H).

Example 50A 1-[4-Amino-2-(piperidin-3-ylamino)-1,3-thiazol-5-yl]ethanonedihydrochloride

Analogously to the preparation of Example 38A, 160 mg (0.15 mmol) oftert-butyl3-[(5-acetyl-4-amino-1,3-thiazol-2-yl)amino]piperidine-1-carboxylate(Example 49A) and 20 ml of hydrochloric acid in dioxane (4M) gave 40 mg(87% of theory) of the product as a solid.

LCMS (method 9): R_(t)=0.81 min. (m/z=241 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=9.22 (s, br, 2H), 8.93 (d, 1H), 7.7 (s, br,1H), 3.96 (m, 1H), 3.35 (d, 1H), 3.12 (d, 1H), 2.86 (m, 2H), 2.05 (s,3H), 1.93-2.05 (m, 1H), 1.8-1.93 (m, 1H), 1.61-1.78 (m, 1H), 1.45-1.6(m, 1H).

Example 51A Methyl2-[(tert-butoxycarbonyl)amino]-6-chloropyridine-3-carboxylate

2.0 g (8.7 mmol) of tert-butyl (6-chloropyridin-2-yl)carbamate (Example24A) were initially charged in 50 ml of THF and cooled to −78° C. 13.7ml (22 mmol) of butyllithium (1.6 M) were added dropwise. After thedropwise addition had ended, the reaction was slowly warmed to −10° C.and kept at −10° C. for 2 h. The mixture was then once more cooled to−78° C., and 870 mg (9.2 mmol) of methyl chloroformate were added. Thereaction solution was warmed to RT over a period of 12 h, and thereaction mixture was then poured into 150 ml of ethyl acetate and 80 mlof hydrochloric acid solution (1N) and stirred for 15 min. The organicphase was separated off, washed with water and saturated sodiumbicarbonate solution, dried over magnesium sulfate and concentrated on arotary evaporator. The reaction mixture was chromatographed on silicagel (mobile phase cyclohexane/ethyl acetate 10:1). This gave 1018 mg(33% of theory) of the product as an oil.

LCMS (method 8): R_(t)=1.25 min. (m/z=187 (M+H-Boc)⁺)

Example 52A Methyl2-[(tert-butoxycarbonyl)amino]-6-({2-[(tert-butoxycarbonyl)amino]ethyl}amino)pyridine-3-carboxylate

Analogously to the preparation of Example 33A, 650 mg (2.3 mmol) ofmethyl 2-[(tert-butoxycarbonyl)amino]-6-chloropyridine-3-carboxylate(Example 51A) and 363 mg (2.3 mmol) of N-Boc-ethylenediamine gave 500 mg(50% of theory) of the product as a solid.

LCMS (method 8): R_(t)=1.32 min. (m/z=411 (M+H)⁺).

Example 53A Methyl 2-amino-6-[(2-aminoethyl)amino]pyridine-3-carboxylatedihydrochloride

Analogously to the preparation of Example 38A, 496 mg (1.2 mmol) ofmethyl2-[(tert-butoxycarbonyl)amino]-6-({2-[(tert-butoxycarbonyl)amino]ethyl}amino)pyridine-3-carboxylate(Example 52A) gave 363 mg (82% of theory) of the product as a solid.

LCMS (method 9): R_(t)=0.75 min. (m/z=212 (M+H-2HCl)⁺).

Example 54A6-[(2-{[6-(2,4-Dichlorophenyl)-2-formylpyrazolo[1,5-a]pyrazin-4-yl]amino}ethyl)amino]pyridine-3-carbonitrile

1.2 g (2.64 mmol) of6-[(2-{[6-(2,4-dichlorophenyl)-2-(hydroxymethyl)pyrazolo[1,5-a]pyrazin-4-yl]amino}ethyl)amino]pyridine-3-carbonitrile(Example 42) were suspended in 80 ml of dichloromethane (shaken withwater beforehand). 80 ml of 1,2-dimethoxyethane and 40 ml of DMF werethen added, and the mixture was cooled to 0° C. 2.46 g (5.81 mmol) ofDess-Martin periodinane were added, ice-bath cooling was removed and themixture was stirred at RT for 2 h. Ethyl acetate was added, and themixture was washed first with a 10% strength sodium thiosulfate solutionand then with saturated aqueous sodium bicarbonate solution. Drying ofthe organic phase with sodium sulfate and removal of the solvent underreduced pressure gave 1.2 g (94% of theory) of the product as a solidwhich was reacted without further purification.

LCMS (method 9): R_(t)=2.50 min. (m/z=452 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=10.1 (s, 1H), 8.38 (d, 1H), 8.33 (s, 1H),8.30 (t, 1H), 7.77 (m, 1H), 7.75 (d, 1H), 7.67 (d, 1H), 7.59 (d, 1H),7.55 (s, 1H), 7.53 (dd, 1H), 6.52 (d, 1H), 3.55-3.7 (m, 4H).

Example 55A6-(2,4-Dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carboxylicacid

500 mg (1.42 mmol) of ethyl6-(2,4-dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carboxylate(Example 11A) were suspended in 60 ml of 1,2-dimethoxyethane, 1M aqueoussodium hydroxide (5.7 ml, 5.7 mmol) was added and the mixture wasstirred at RT for 3 h. The reaction mixture was poured into water andacidified to pH 2 using 2M aqueous hydrochloric acid. The precipitatewas filtered off and washed with water. Drying gave 430 mg (93% oftheory) of the product as a solid.

LCMS (method 6): R_(t)=1.31 min. (m/z=324 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=13.29 (br, 1H), 11.80 (s, 1H), 7.94 (s,1H), 7.84 (d, 1H), 7.66 (d, 1H), 7.59 (dd, 1H), 7.48 (s, 1H).

Example 56A6-(2,4-Dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carboxamide

360 mg (1.11 mmol) of6-(2,4-dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carboxylicacid (Example 55A) were initially charged in dichloromethane (30 ml) andDMF (10 ml), and 320 mg (1.67 mmol) of EDC, 225 mg (1.67 mg) HOBt and407 mg (3.33 mmol) of DMAP were added, followed by 0.5M ammonia indioxane (2.44 ml, 1.22 mmol). The reaction mixture was stirred at RT for20 h, the solvent was concentrated, the residue was poured into water(25 ml) and the precipitate was filtered off and washed with water andacetonitrile and dried. Drying gave 260 mg (72% of theory) of theproduct as a solid.

LCMS (method 3): R_(t)=1.69 min. (m/z=323 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=11.77 (br, 1H), 7.86 (s, 1H), 7.84 (m, 2H),7.65 (d, 1H), 7.59 (m, 2H), 7.47 (s, 1H).

Example 57A4-Chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carbonitrile

Analogously to the preparation of Example 16A, 260 mg (0.81 mmol) of6-(2,4-dichlorophenyl)-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2-carboxamide(Example 56A) gave, by reaction with phosphoryl chloride, 230 mg (77% oftheory) of the product as a solid.

LCMS (method 3): R_(t)=2.86 min. (m/z=323 (M+H)⁺).

WORKING EXAMPLES Example 16-[(2-{[6-(2,4-Dichlorophenyl)imidazo[1,2-a]pyrazin-8-yl]amino}ethyl)amino]nicotinonitrile

108 mg (0.3625 mmol) of the8-chloro-6-(2,4-dichlorophenyl)imidazo[1,2-a]pyrazine (Example 5A) wereinitially charged in 3 ml of DMSO, and 150 mg (0.543 mmol) of6-[(2-aminoethyl)amino]nicotinonitrile (Example 2A) and 0.63 ml (3.62mmol) of N,N-diisopropylethylamine were added. The mixture was heated at120° C. for 12 h. Purification by chromatography on silica gel 60(mobile phase: dichloromethane/methanol 100:1) gave 12 mg (7% of theory)of the product.

LCMS (method 6): R_(t)=1.97 min. (m/z=424 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.35 (d, 1H), 8.09 (s, 1H), 7.94 (s, 1H),7.74 (s, br, 1H), 7.70 (d, 2H), 7.63 (d, 1H), 7.56 (d, 2H), 7.49 (dd,1H), 6.53 (s, br, 1H), 3.66 (t, 2H), 3.58 (t, br, 2H).

Example 24-Amino-2-[(2-{[6-(2,4-dichlorophenyl)imidazo[1,2-a]pyrazin-8-yl]amino}ethyl)amino]-1,3-thiazole-5-carbonitriletrifluoroacetate

Under argon, 80 mg (0.2 mmol) of4-amino-2-({2-[(6-bromoimidazo[1,2-a]pyrazin-8-yl)amino]ethyl}amino)-1,3-thiazole-5-carbonitrile(Example 9A) were initially charged in 4.5 ml of dioxane and 1.3 ml ofsaturated sodium carbonate solution, and 48 mg (0.25 mmol) of2,4-dichlorobenzeneboronic acid and 22 mg (0.02 mmol) oftetrakis(triphenylphosphine)palladium(0) were added. The mixture washeated in a microwave at 160° C. for 1 h. The reaction mixture wasconcentrated on a rotary evaporator. This gave, after purification bypreparative HPLC, 35 mg (33% of theory) of the product as a solid.

LCMS (method 8): R_(t)=1.16 min. (m/z=444 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.48 (t, 1H), 8.15 (s, 1H), 7.99 (s, 1H),7.9 (s, broad, 1H), 7.72 (d, 1H), 7.65 (d, 1H), 7.62 (s, 1H), 7.51 (dd,1H), 3.67 (q, 2H), 3.49 (q, 2H).

Example 3 Ethyl4-({2-[(5-cyanopyridin-2-yl)amino]ethyl}amino)-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]-pyrazine-2-carboxylate

1 g (5 mmol) of ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 12A) was initially charged in 15 ml of dry DMSO, and 1.55 g(4.22 mmol) of 6-[(2-aminoethyl)amino]nicotinonitrile dihydrochloride(Example 2A) and 5.8 ml (33.5 mmol) of N,N-diisopropylethylamine wereadded and the mixture was heated in a microwave at 150° C. for 30 min.Ethyl acetate and 10% strength citric acid were added, and the reactionmixture was extracted. The organic phase was washed with sodium chloridesolution and dried over magnesium sulfate. This gave, afterconcentration, 1.4 g (70% of theory) of the product as a solid.

LCMS (method 6): R_(t)=2.31 min. (m/z=496 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.38 (d, 1H), 8.3 (s, 1H), 8.15 (t, 1H),7.73 (d, 1H), 7.66 (d, 2H), 7.59 (dd, 1H), 7.57 (s, 1H), 7.52 (dd, 1H),6.52 (d, 1H), 4.35 (t, 2H), 3.7-3.55 (m, 4H), 1.34 (s, 3H).

Example 44-({2-[(5-Cyanopyridin-2-yl)amino]ethyl}amino)-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylicacid hydrochloride

80 mg (0.16 mmol) of ester (Example 3) were dissolved in 5 ml of1,2-dimethoxyethane, and 2.5 ml of water and 0.43 ml (0.4 mmol) of 1Nsodium hydroxide solution were added. The solution was stirred at RT for2 h. 20 ml of dilute hydrochloric acid (pH 3) were initially charged,and the reaction solution was slowly added dropwise. The mixture wasstirred for another 30 min, and the product was filtered off and washedwith diethyl ether. This gave, after drying under high vacuum, 70 mg(84% of theory) of the product as a solid.

LCMS (method 3): R_(t)=2.59 min. (m/z=468 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.38 (d, 1H), 8.26 (s, 1H), 8.17 (t, 1H),7.77 (s, broad, 1H), 7.73 (d, 1H), 7.66 (d, 1H), 7.59 (d, 1H), 7.52 (dd,1H), 7.51 (s, 1H), 6.53 (d, 1H), 3.7-3.56 (m, 4H).

General Description for Amide Coupling Experiments:

0.16 mmol of acid (Example 4) is initially charged in 2 ml of DMF, and0.17 mmol of HATU, 0.48 mmol of N,N-diisopropylethylamine and 0.21 mmolof amine are added in succession. The mixture is stirred at RT for 12 h.Purification by preparative HPLC gives 60%-95% of theory of the productas a solid.

The compounds below were prepared according to the general descriptionfor amide coupling experiments.

Ex. Structure Characterization 5

LC/MS (method 8): R_(t) = 0.89 min, (m/z = 593 (M + H)⁺) ¹H-NMR (400MHz, DMSO-d₆): δ = 10.81 (s, 1H), 8.38 (d, 1H), 8.28 (s, 1H), 8.22 (s,1H), 7.91 (s, 1H), 7.74 (d, 1H), 7.66 (d, 1H), 7.61 (d, 1H), 7.53 (dd,1H), 7.45 (s, 1H), 6.56 (d, 1H), 4.89 (d, 1H), 4.61 (d, 1H), 3.65 (m,6H), 3.48 (q, 3H), 3.27 (t, 1H), 3.12 (m, 4H), 2.79 (d, 3H). 6

LC/MS (method 6): R_(t) = 1.60 min, (m/z = 551 (M + H)⁺) ¹H-NMR (400MHz, DMSO-d₆): δ = 8.38 (d, 1H), 8.28 (s, 1H), 8.22 (s, 1H), 7.91 (s,1H), 7.74 (d, 1H), 7.66 (d, 1H), 7.61 (d, 1H), 7.53 (dd, 1H), 7.45 (s,1H), 6.56 (d, 1H), 3.67 (m, 8H), 3.38 (s, broad, 4H), 2.38 (s, 3H). 7

LC/MS (method 8): R_(t) = 1.28 min, (m/z = 537 (M + H)⁺) ¹H-NMR (400MHz, DMSO-d₆): δ = 8.39 (d, 1H), 8.25 (s, broad, 1H), 8.23 (s, 1H), 7.73(d, 1H), 7.66 (d, 2H), 7.63 (dd, 1H), 7.52 (d, 1H), 7.41 (s, 1H), 6.6(d, 1H), 3.73 (m, broad, 8H), 3.63 (s, broad, 4H). 8

LC/MS (method 8): R_(t) = 0.98 min, (m/z = 538 (M + H)⁺) ¹H-NMR (400MHz, DMSO-d₆): δ = 10.09 (s, 1H), 8.8 (t, 1H), 8.39 (d, 2H), 8.19 (s,1H), 7.75 (d, 1H), 7.7 (d, 1H), 7.63 (dd, 1H), 7.54 (m, 2H), 6.62 (d,1H), 3.73-3.58 (m, 6H), 3.27 (q, 2H), 2.8 (d, 6H).

Example 96-({2-[(5,7-Dimethyl-2-phenylimidazo[5,1-F][1,2,4]triazin-4-yl)-amino]ethyl}amino)nicotinonitrile

183 mg (0.71 mmol) of the4-chloro-5,7-dimethyl-2-phenylimidazo[5,1-f][1,2,4]triazine (Example16A) were initially charged in 4 ml of DMSO, and 200 mg (0.85 mmol) of6-[(2-aminoethyl)-amino]nicotinonitrile (Example 2A) and 1.23 ml (7mmol) of N,N-diisopropylethylamine were added and the mixture was heatedat 150° C. for 12 h. Purification by chromatography on silica gel(mobile phase: dichloromethane/methanol 100:1) gave 10 mg (4% of theory)of the product.

LCMS (method 6): R_(t)=1.40 min. (m/z=385 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.43 (s, 1H), 8.12 (d, 1H), 7.99 (d, 1H),7.82 (t, 1H), 7.59 (t, 2H), 7.55 (q, 1H), 7.46 (m, 2H), 6.50 (d, 1H),3.86 (q, 2H), 3.67 (s, broad, 2H), 2.5 (s, 6H).

Example 104-Amino-2-{[2-({6-[4-(trifluoromethyl)phenyl]imidazo[1,2-a]pyrazin-8-yl}amino)ethyl]amino}-1,3-thiazole-5-carbonitriletrifluoroacetate

Under argon, 80 mg (0.192 mmol) of4-amino-2-({2-[(6-bromoimidazo[1,2-a]pyrazin-8-yl)amino]-ethyl}amino)-1,3-thiazole-5-carbonitrile(Example 9A) were initially charged in 4.5 ml of dioxane and 1.3 ml ofsaturated sodium carbonate solution, and 47 mg (0.25 mmol) of4-(trifluoromethyl)-phenyl]boronic acid and 22 mg (0.019 mmol) oftetrakis(triphenylphosphine)palladium(0) were added. The mixture washeated at 120° C. for 15 h. The reaction mixture was concentrated on arotary evaporator. This gave, after purification by preparative HPLC, 5mg (5% of theory) of the product as a solid.

LCMS (method 8): R_(t)=1.16 min. (m/z=445 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.59 (s, 1H), 8.53 (t, 1H), 8.13 (d, 2H),7.93 (s, 1H), 7.86 (t, 1H), 7.79 (d, 2H), 7.58 (s, 1H), 6.74 (s, broad,2H), 3.80 (q, 2H), 3.54 (q, 2H).

Example 11 Ethyl4-({2-[(4-amino-5-cyano-1,3-thiazol-2-yl)-amino]ethyl}amino)-6-(2,4-dichlorophenyl)-pyrazolo[1,5-a]pyrazine-2-carboxylate

Analogously to the procedure described for Example 3, 300 mg (0.769mmol) of ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 12A) gave, by reaction with 239 mg (0.92 mmol) of4-amino-2-[(2-aminoethyl)amino]-1,3-thiazole-5-carbonitriledihydrochloride (Example 8A) and purification by preparative HPLC, 110mg (26% of theory) of the product as a solid.

LCMS (method 3): R_(t)=2.58 min. (m/z=517 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.49 (t, 1H), 8.32 (s, 1H), 8.17 (t, 1H),7.74 (d, 1H), 7.66 (d, 1H), 7.58 (s, 1H), 7.53 (dd, 1H), 6.70 (s, br,2H), 4.35 (q, 2H), 3.66 (dd, 2H), 3.52 (dd, 2H), 1.34 (t, 3H).

Example 12 Ethyl4-{3-[(4-amino-5-cyano-1,3-thiazol-2-yl)-amino]piperidin-1-yl}-6-(2,4-dichlorophenyl)-pyrazolo[1,5-a]pyrazine-2-carboxylate

Analogously to the procedure described for Example 3, 52.1 mg (0.133mmol) of ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 12A) gave, by reaction with 132 mg (0.2 mmol) of4-amino-2-(piperidin-3-ylamino)-1,3-thiazole-5-carbonitriledihydrochloride (Example 47A) and purification by preparative HPLC, 52mg (70% of theory) of the product as a solid.

LCMS (method 8): R_(t)=1.47 min. (m/z=557 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.8 (s, 2H), 7.74 (d, 1H), 7.72 (d, 1H),7.6 (s, 1H), 7.54 (dd, 1H), 6.72 (s, 2H), 4.3-4.44 (m, 3H), 4.08 (d,1H), 3.86 (s, br, 1H), 3.4-3.5 (m, 2H), 2.04 (m, 1H), 1.90 (m, 1H), 1.65(t, 2H), 1.35 (t, 3H).

Example 13 Ethyl4-{3-[(6-amino-5-nitropyridin-2-yl)amino]piperidin-1-yl}-6-(2,4-dichlorophenyl)pyrazolo-[1,5-a]pyrazine-2-carboxylatetrifluoroacetate

Analogously to the procedure described for Example 3, 70 mg (0.179 mmol)of ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 12A) gave, by reaction with 83 mg (0.27 mmol) of3-nitro-N⁶-(piperidin-3-yl)pyridine-2,6-diamine dihydrochloride (Example42A) and purification by preparative HPLC, 83 mg (67% of theory) of theproduct as a solid.

LCMS (method 6): R_(t)=2.56 min. (m/z=571 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.48 (s, 1H), 8.14 (s, br, 1H), 7.95 (d,1H), 7.90 (d, 1H), 7.72 (m, 3H), 7.51 (dd, 1H), 7.39 (s, 1H), 5.92 (d,1H), 4.32 (q, 2H), 4.19 (d, 1H), 3.98 (m, 1H), 3.64 (m, 2H), 2.0 (m,2H), 1.69 (m, 2H), 1.30 (t, 3H).

Example 14 Ethyl4-{3-[(6-amino-5-nitropyridin-2-yl)amino]piperidin-1-yl}-6-(2,4-dichlorophenyl)pyrazolo-[1,5-a]pyrazine-2-carboxylatetrifluoroacetate

Analogously to the procedure described for Example 3, 70 mg (0.179 mmol)of ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 12A) gave, by reaction with 83 mg (0.27 mmol) of1-{2-amino-6-[(2-aminoethyl)amino]pyridin-3-yl}-2,2,2-trifluoroethanonehydrochloride (Example 34A) and purification by preparative HPLC (mobilephase: acetonitrile/water gradient with 0.1% trifluoroacetic acid), 83mg (67% of theory) of the product as a solid.

LCMS (method 6): R_(t)=2.56 min. (m/z=571 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.48 (s, 1H), 8.14 (s, br, 1H), 7.95 (d,1H), 7.90 (d, 1H), 7.72 (m, 3H), 7.51 (dd, 1H), 7.39 (s, 1H), 5.92 (d,1H), 4.32 (q, 2H), 4.19 (d, 1H), 3.98 (m, 1H), 3.64 (m, 2H), 2.0 (m,2H), 1.69 (m, 2H), 1.30 (t, 3H).

Example 15 Ethyl4-(3-{[4-amino-5-(methoxycarbonyl)-1,3-thiazol-2-yl]amino}piperidin-1-yl)-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate

Analogously to the procedure described for Example 3, 63 mg (0.162 mmol)of ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 12A) gave, by reaction with 89.9 mg (0.243 mmol) of methyl4-amino-2-(piperidin-3-ylamino)-1,3-thiazole-5-carboxylatedihydrochloride (Example 45A) and purification by preparative HPLC, 68mg (69% of theory) of the product as a solid.

LCMS (method 6): R_(t)=2.58 min. (m/z=590 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.50 (s, 1H), 8.47 (d, 1H), 7.72 (d, 2H),7.65 (s, 1H), 7.52 (dd, 1H), 6.81 (s, br, 2H), 4.37 (m, 3H), 4.13 (dt,1H), 3.87 (m, 1H), 3.61 (s, 3H), 2.06 (m, 1H), 1.92 (m, 1H), 1.65 (m,2H), 1.34 (t, 3H).

Example 16 Ethyl4-{3-[(6-amino-5-nitropyridin-2-yl)amino]piperidin-1-yl}-6-(2,4-dichlorophenyl)pyrazolo-[1,5-a]pyrazine-2-carboxylatetrifluoroacetate

Analogously to the procedure described for Example 3, 70 mg (0.179 mmol)of ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 12A) gave, by reaction with mg (0.27 mmol) of methyl2-amino-6-[(2-aminoethyl)amino]pyridine-3-carboxylate dihydrochloride(Example 53A) and purification by preparative HPLC (mobile phase:acetonitrile/water gradient with 0.1% trifluoroacetic acid), 83 mg (67%of theory) of the product as a solid.

LCMS (method 6): R_(t)=2.56 min. (m/z=571 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.48 (s, 1H), 8.14 (s, br, 1H), 7.95 (d,1H), 7.90 (d, 1H), 7.72 (m, 3H), 7.51 (dd, 1H), 7.39 (s, 1H), 5.92 (d,1H), 4.32 (q, 2H), 4.19 (d, 1H), 3.98 (m, 1H), 3.64 (m, 2H), 2.0 (m,2H), 1.69 (m, 2H), 1.30 (t, 3H).

Example 17 Ethyl4-{3-[(5-acetyl-4-amino-1,3-thiazol-2-yl)amino]piperidin-1-yl}-6-(2,4-dichlorophenyl)-pyrazolo[1,5-a]pyrazine-2-carboxylatetrifluoroacetate

Analogously to the procedure described for Example 3, 61 mg (0.155 mmol)of ethyl4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 12A) gave, by reaction with mg (0.233 mmol) of1-[4-amino-2-(piperidin-3-ylamino)-1,3-thiazol-5-yl]ethanonedihydrochloride (Example 50A) and purification by preparative HPLC(mobile phase: acetonitrile/water gradient with 0.1% trifluoroaceticacid), 63 mg (59% of theory) of the product as a solid.

LCMS (method 6): R_(t)=2.40 min. (m/z=574 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.65 (d, 1H), 8.51 (s, 1H), 7.74 (d, 1H),7.71 (d, 1H), 7.64 (s, 1H), 7.52 (dd, 1H), 4.31-4.43 (m, 3H), 4.13 (d,2H), 3.85 (s, br, 1H), 3.44 (m, 2H), 2.06 (m, 1H), 1.98 (s, 3H), 1.92(m, 1H), 1.6-1.72 (m, 2H), 1.34 (t, 3H).

Analogously to the procedure described for Example 4, the esters inquestion were converted by hydrolysis with lithium hydroxide or sodiumhydroxide solution in the corresponding acids.

Ex. Structure Characterization 18

LC/MS (method 8): R_(t) = 1.11 min, (m/z = 489 (M + H)⁺) 19

LC/MS (method 6): R_(t) = 2.10 min, (m/z = 529 (M + H)⁺) 20

LC/MS (method 6): R_(t) = 1.98 min, (m/z = 468 (M + H)⁺) ¹H-NMR (400MHz, DMSO-d₆): δ = 8.79 (s, 1H), 8.45 (s, 1H), 8.24 (d, 2H), 8.18 (t,1H), 7.84 (br, 1H), 7.79 (d, 2H), 7.63 (d, 1H), 7.51 (s, 1H), 6.53 (m,1H), 3.79 (m, 2H), 3.66 (m, 2H).

The amides below were prepared analogously to the general descriptionfor amide coupling experiments (cf. Example 5) from the correspondingcarboxylic acids.

Ex. Structure Characterization 21

LC/MS (method 3): R_(t) = 2.69 min, (m/z = 602 (M + H)⁺) 22

LC/MS (method 6): R_(t) = 1.38 min, (m/z = 592 (M + H)⁺) 23

LC/MS (method 3): R_(t) = 2.95 min, (m/z = 581 (M + H)⁺) 24

LC/MS (method 6): R_(t) = 1.74 min, (m/z = 546 (M + H)⁺) 25

LC/MS (method 3): R_(t) = 2.43 min, (m/z = 525 (M + H)⁺) 26

LC/MS (method 8): R_(t) = 1.22 min, (m/z = 539 (M + H)⁺) 27

LC/MS (method 6): R_(t) = 2.17 min, (m/z = 567 (M + H)⁺) 28

LC/MS (method 9): R_(t) = 2.20 min, (m/z = 558 (M + H)⁺) 29

LC/MS (method 3): R_(t) = 1.57 min, (m/z = 559 (M + H)⁺) 30

LC/MS (method 3): R_(t) = 1.63 min, (m/z = 580 (M + H)⁺) 31

LC/MS (method 6): R_(t) = 1.24 min, (m/z = 614 (M + H)⁺) 32

LC/MS (method 8): R_(t) = 1.22 min, (m/z = 467 (M + H)⁺) 33

LC/MS (method 3): R_(t) = 1.57 min, (m/z = 571 (M + H)⁺)

Example 341-{2-Amino-6-[(2-{[6-(2,4-dichlorophenyl)-2-(morpholin-4-ylmethyl)pyrazolo[1,5-a]pyrazin-4-yl]amino}ethyl)amino]pyridin-3-yl}-2,2,2-trifluoroethanonetrifluoroacetate

Analogously to the procedure described for Example 1, 60 mg (0.151 mmol)of4-chloro-6-(2,4-dichlorophenyl)-2-(morpholin-4-ylmethyl)pyrazolo[1,5-a]pyrazine(Example 22A) gave, by reaction with 53 mg (0.181 mmol) of1-{2-amino-6-[(2-aminoethyl)amino]pyridin-3-yl}-2,2,2-trifluoroethanonehydrochloride (Example 34A) and purification by preparative HPLC, 62 mg(57% of theory) of the product as a solid.

LCMS (method 3): R_(t)=1.84 min. (m/z=609 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=10.41 (s, br, 1H), 8.54 (s, 1H), 8.26 (s,1H), 8.10 (s, 2H), 7.73 (d, 1H), 7.68 (d, 1H), 7.54 (s, br, 1H), 7.48(dd, 1H), 7.14 (s, 1H), 5.91 (d, 1H), 4.59 (s, 2H), 3.96 (m, 2H),3.51-3.75 (m, 6H), 3.39 (m, 2H), 3.19 (m, 2H).

Analogously to the procedure described for the preparation of Example 1,the products below were synthesized from the corresponding chlorides byreaction with the appropriate amines.

Ex. Structure Characterization 35

LC/MS (method 3): R_(t) = 1.56 min, (m/z = 557 (M + H)⁺) 36

LC/MS (method 3): R_(t) = 1.74 min, (m/z = 578 (M + H)⁺) 37

LC/MS (method 8): R_(t) = 0.98 min, (m/z = 571 (M + H)⁺) 38

LC/MS (method 6): R_(t) = 1.73 min, (m/z = 649 (M + H)⁺) 39

LC/MS (method 3): R_(t) = 1.84 min, (m/z = 563 (M + H)⁺) 40

LC/MS (method 3): R_(t) = 1.59 min, (m/z = 538 (M + H)⁺)

Example 414-Amino-2-[(2-{[6-(2,4-dichlorophenyl)-2-(hydroxymethyl)pyrazolo[1,5-a]pyrazin-4-yl]amino}-ethyl)amino]-1,3-thiazole-5-carbonitrile

100 mg (0.184 mmol) of ethyl4-({2-[(4-amino-5-cyano-1,3-thiazol-2-yl)amino]ethyl}amino)-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 11) were dissolved in 10 ml of THF, and 0.276 ml (0.276 mmol)of a solution of lithium aluminum hydride (1 mol/l) in THF was addeddropwise at RT. The mixture was stirred for 2 h, and after completeconversion first methanol was added, and then the pH was adjusted topH=5 using dilute hydrochloric acid. The mixture was extractedrepeatedly with ethyl acetate. Removal of the solvent and purificationby preparative HPLC gave 61 mg (64% of theory) of the product as asolid.

LCMS (method 8): R_(t)=1.10 min. (m/z=475 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.50 (t, 1H), 8.15 (s, 1H), 7.91 (t, 1H),7.70 (d, 1H), 7.68 (d, 1H), 7.51 (dd, 1H), 6.95 (s, 2H), 6.69 (s, br,2H), 4.62 (s, 2H), 3.64 (dd, 2H), 3.51 (m, 2H).

Example 424-Amino-2-[(2-{[6-(2,4-dichlorophenyl)-2-(hydroxymethyl)pyrazolo[1,5-a]pyrazin-4-yl]amino}-ethyl)amino]-1,3-thiazole-5-carbonitrile

100 mg (0.184 mmol) of ethyl4-({2-[(4-amino-5-cyano-1,3-thiazol-2-yl)amino]ethyl}amino)-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carboxylate(Example 3) were dissolved in 10 ml of THF, and 0.276 ml (0.276 mmol) ofa solution of lithium aluminum hydride (1 mol/l) in THF was addeddropwise at RT. The mixture was stirred for 2 h, and after completeconversion first methanol was added, and then the pH was adjusted topH=5 using dilute hydrochloric acid. The mixture was extractedrepeatedly with ethyl acetate. Removal of the solvent and purificationby preparative HPLC gave 61 mg (64% of theory) of the product as asolid.

LCMS (method 8): R_(t)=1.10 min. (m/z=475 (M+H)⁺)

¹H-NMR (400 MHz, DMSO-d₆): δ=8.50 (t, 1H), 8.15 (s, 1H), 7.91 (t, 1H),7.70 (d, 1H), 7.68 (d, 1H), 7.51 (dd, 1H), 6.95 (s, 2H), 6.69 (s, br,2H), 4.62 (s, 2H), 3.64 (dd, 2H), 3.51 (m, 2H).

Analogously to the procedure described for Example 2, the products belowwere obtained from6-({2-[(6-bromoimidazo[1,2-a]pyrazin-8-yl)amino]ethyl}amino)pyridine-3-carbonitrile(Example 23A) by palladium-catalyzed reaction with the correspondingboronic acids.

Ex. Structure Characterization 43

LC/MS (method 8): R_(t) = 1.32 min, (m/z = 398 (M + H)⁺) 44

LC/MS (method 10): R_(t) = 2.02 min, (m/z = 392 (M + H)⁺) 45

LC/MS (method 11): R_(t) = 2.04 min, (m/z = 392 (M + H)⁺) 46

LC/MS (method 10): R_(t) = 1.43 min, (m/z = 399 (M + H)⁺) 47

LC/MS (method 11): R_(t) = 2.16 min, (m/z = 440 (M + H)⁺) 48

LC/MS (method 11): R_(t) = 1.66 min, (m/z = 384 (M + H)⁺) 49

LC/MS (method 11): R_(t) = 1.92 min, (m/z = 374 (M + H)⁺) 50

LC/MS (method 10): R_(t) = 2.11 min, (m/z = 410 (M + H)⁺) 51

LC/MS (method 11): R_(t) = 2.08 min, (m/z = 390 (M + H)⁺) 52

LC/MS (method 10): R_(t) = 1.82 min, (m/z = 362 (M + H)⁺)

Example 536-[(2-{[6-(2,4-Dichlorophenyl)-2-(morpholin-4-ylmethyl)pyrazolo[1,5-a]pyrazin-4-yl]amino}ethyl)amino]nicotinonitrilehydrochloride

30 mg (0.066 mmol) of6-[(2-{[6-(2,4-dichlorophenyl)-2-formylpyrazolo[1,5-a]pyrazin-4-yl]amino}ethyl)amino]pyridine-3-carbonitrile(Example 54A) were dissolved in 1 ml of methanol, and 11.6 mg (0.133mmol) of morpholine, 4 Å molecular sieve and 11.9 mg (0.199 mmol) ofacetic acid were added. Finally, 8.3 mg (0.133 mmol) of sodiumcyanoborohydride were added and the mixture was stirred at RT for 1 h.The crude mixture was acidified with 2N hydrochloric acid and purifiedby preparative HPLC. Lyophilization gave 25 mg (58% of theory) of theproduct as a solid.

LCMS (method 3): R_(t)=1.74 min. (m/z=523 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=11.0 (br, 1H), 8.39 (d, 1H), 8.27 (m, 1H),8.23 (s, 1H), 7.87 (br, 1H), 7.74 (d, 1H), 7.67 (d, 1H), 7.61 (m, 1H),7.54 (dd, 1H), 7.23 (s, 1H), 6.57 (m, 1H), 4.57 (s, 2H), 3.95 (m, 4H),3.62 (m, 4H), 3.49 (m, 2H), 3.18 (m, 2H).

Analogously to the procedure described for the preparation of Example53, the products below were obtained by reductive amination startingwith6-[(2-{[6-(2,4-dichlorophenyl)-2-formylpyrazolo[1,5-a]pyrazin-4-yl]amino}ethyl)amino]pyridine-3-carbonitrile(Example 54A) or the corresponding amines.

Ex. Structure Characterization 54

LCMS (method 9): R_(t) = 1.72 min. (m/z = 536 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 8.40 (d, 1H), 8.23 (s, 1H), 8.18 (m, 1H), 7.86 (br,1H), 7.74 (d, 1H), 7.66 (d, 1H), 7.61 (m, 1H), 7.52 (dd, 1H), 7.13 (m,1H), 6.50 (m, 1H), 3.50 (m, 2H), 3.20 (m, 2H), 2.81 (s, 3H). Othersignals overlap with the signal for water. 55

LCMS (method 9): R_(t) = 1.50 min. (m/z = 524 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 10.74 (br, 1H), 9.83 (br, 1H), 8.42 (d, 1H), 8.29 (m,1H), 8.23 (s, 1H), 7.94 br, 1H), 7.75 (d, 1H), 7.69 (d, 1H), 7.54 (dd,1H), 7.20 (s, 1H), 6.56 (m, 1H), 4.45 (m, 2H), 3.66 (m, 4H), 3.47 (m,4H), 2.85 (d, 3H). 56

LCMS (method 9): R_(t) = 1.71 min. (m/z = 522 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 11.75 (br, 1H), 9.25 (br, 2H), 8.40 (d, 1H), 8.23 (s,2H), 7.84 (br, 1H), 7.74 (d, 1H), 7.66 (d, 1H), 7.60 (d, 1H), 7.53 (dd,1H), 7.21 (s, 1H), 6.57 (m, 1H), 4.50 (br s, 2H). Other signals overlapwith the signal for water. 57

LCMS (method 9): R_(t) = 1.83 min. (m/z = 509 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 10.25 (br, 1H), 8.39 (d, 1H), 8.24 (s, 1H), 8.22 (m,1H), 7.87 (br, 1H), 7.74 (d, 1H), 7.68 (d, 1H), 7.61 (br d, 1H), 7.53(dd, 1H), 7.24 (s, 1H), 6.56 (m, 1H), 4.54 (m, 2H), 3.65 (m, 4H), 3.13(m, 4H), 1.31 (t, 6H). 58

LCMS (method 9): R_(t) = 1.79 min. (m/z = 493 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 8.37 (d, 1H), 8.13 (s, 1H), 7.85 (m, 1H), 7.76 (br,1H), 7.72 (d, 1H), 7.68 (d, 1H), 7.59 (d, 1H), 7.51 (dd, 1H), 6.90 (s,1H), 6.54 (m, 1H), 3.89 (s, 2H), 3.64 (m, 4H), 2.15 (m, 1H), 0.37 (m,2H), 0.28 (m, 2H). 59

LCMS (method 9): R_(t) = 1.94 min. (m/z = 578 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 11.25 (br, 1H), 8.39 (d, 1H), 8.27 (m, 1H), 8.23 (s,1H), 7.89 (br, 1H), 7.75 (d, 1H), 7.68 (d, 1H), 7.62 (d, 1H), 7.54 (dd,1H), 7.24 (s, 1H), 6.57 (m, 1H), 4.58 (m, 2H), 3.68 (m, 8H), 3.45 (m,4H), 1.36 (s, 9H). 60

LCMS (method 8): R_(t) = 1.04 min. (m/z = 536 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 8.73 (s, 1H), 8.45 (s, 1H), 8.25 (d, 2H), 8.16 (br,1H), 7.91 (br, 1H), 7.77 (d, 2H), 7.64 (m, 1H), 7.14 (m, 1H), 6.56 (m,1H), 2.81 (s, 3H). Other signals overlap with the signal for water. 61

LCMS (method 8): R_(t) = 0.95 min. (m/z = 524 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 10.61 (br, 1H), 9.75 (br, 2H), 8.70 (s, 1H), 8.45 (m,1H), 8.26 (d, 2H), 8.22 (m, 1H), 7.90 7.79 (d, 2H), 7.64 (d, 1H), 7.17(s, 1H), 6.54 (m, 1H), 4.47 (m, 2H), 3.79 (m, 2H), 2.86 (d, 6H). Othersignals overlap with the signal for water. 62

LCMS (method 9): R_(t) = 1.84 min. (m/z = 511 (M + H)⁺). ¹H-NMR (400MHz, DMSO-d₆): δ = 9.67 (br, 2H), 8.409 (d, 1H), 8.25 (m, 1H), 8.23 (s,1H), 7.92 (br, 1H), 7.74 (d, 1H), 7.68 (d, 1H), 7.63 (d, 1H), 7.54 (dd,2H), 7.16 (s, 1H), 6.57 (m, 1H), 4.41 (m, 2H), 3.96 (m, 2H), 3.64 (m,4H).

Example 634-({2-[(4-Amino-5-cyano-1,3-thiazol-2-yl)-amino]ethyl}amino)-6-(2,4-dichlorophenyl)pyrazolo-[1,5-a]pyrazine-2-carbonitrile

Analogously to the procedure described for Example 3, 60 mg (0.19 mmol)of 4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carbonitrile(Example 57A) gave, by reaction with 61.1 mg (0.28 mmol) of4-amino-2-[(2-aminoethyl)amino]-1,3-thiazole-5-carbonitriledihydrochloride (Example 8A) and purification by preparative HPLC, 24 mg(28% of theory) of the product as a solid.

LCMS (method 3): R_(t)=2.63 min. (m/z=470 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=8.50 (t, 1H), 8.37 (s, 1H), 8.35 (t, 1H),7.76 (d, 1H), 7.66 (m, 2H), 7.54 (dd, 1H), 6.69 (br, 2H), 3.78 (m, 2H),3.53 (m, 2H).

Example 644-({2-[(5-Cyanopyridin-2-yl)amino]ethyl}amino)-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carbonitrile

Analogously to the procedure described for Example 3, 60 mg (0.19 mmol)of 4-chloro-6-(2,4-dichlorophenyl)pyrazolo[1,5-a]pyrazine-2-carbonitrile(Example 57A) gave, by reaction with 61.1 mg (0.28 mmol) of6-[(2-aminoethyl)amino]pyridine-3-carbonitrile dihydrochloride (Example2A) and purification by preparative HPLC, 42 mg (50% of theory) of theproduct as a solid.

LCMS (method 3): R_(t)=2.82 min. (m/z=449 (M+H)⁺).

¹H-NMR (400 MHz, DMSO-d₆): δ=8.34 (m, 3H), 7.76 (m, 2H), 7.67 (m, 2H),7.60 (dd, 1H), 7.54 (dd, 1H), 6.51 (m, 1H), 3.69-3.57 (m, 4H).

B) ASSESSMENT OF THE PHYSIOLOGICAL ACTIVITY

The suitability of the compounds according to the invention for treatinghematological disorders can be shown in the following assay systems:

In Vitro Assay

The inhibitory activity of active substances is determined in abiochemical assay. The ingredients required for this purpose are mixedin a black 384-well microtiter plate with transparent base (fromGreiner, catalog number 781092). The requirements in this connection foreach well of the 384-well microtiter plate are 5 nM GSK3β (from Upstate,catalog number xy), 40 μM GSK3β substrate GSM (sequenceH-RRRPASVPPSPSLSRHS-(pS)-HQRR, from Upstate, catalog number 2-533), 30μM nicotinamide adenine dinucleotide NADH (Roche Diagnostics, catalognumber 10107735), 50 μM adenosine triphosphate ATP (from Sigma, catalognumber A7966) and 2 mM phosphoenolpyruvate (from Roche, catalog number128112). The required reaction buffer in which the biochemical reactiontakes place consists of 50 mM Trizma hydrochloride Tris-HCl pH: 7.5(from Sigma, catalog number T3253), 5 mM magnesium chloride MgCl₂ (fromSigma, catalog number M8266), 0.2 mM DL-dithiothreitol DTT (from Sigma,catalog number D9779), 2 mM ethylenediaminetetraacetic acid EDTA (fromSigma, catalog number E6758), 0.01% Triton X-100 (from Sigma, catalognumber T8787) and 0.05% bovine serum albumin BSA (from Sigma, catalognumber B4287).

Active substances are dissolved in dimethyl sulfoxide DMSO (from Sigma,catalog number D8418) in a concentration of 10 mM. Active substances areadded in serial concentrations of 10 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001μM, 0.0001 μM, 0.00001 μM, 0.000001 μM to the mixtures of thebiochemical reaction. As control, dimethyl sulfoxide is added instead ofsubstance in a final concentration of 0.1%.

The reaction is incubated at 30° C. for 2 hours and then the resultingfluorescence is measured in a Tecan Safire-XFLUOR4 instrument, versionV4.50 (serial number 12901300283) with the specifications: measurementmode—fluorescence measured from below, extinction wavelength 340 nm,emission wavelength 465 nm, slit width extinction 5 nm, slit widthemission 5 nm, gain mode 120, delay 0 μs, number of light flashes permeasurement 3, and an integration time of 40 μs.

The GSK3β activity is measured in fluorescence units, with the values ofuninhibited kinase being set equal to 100% and those of completelyinhibited kinase being set equal to 0%. The activity of the activesubstances is calculated in relation to these 0% and 100%.

Table A shows representative in vitro activity data for the compoundsaccording to the invention:

TABLE A Example No. IC₅₀ [nM] 2 16 4 21 5 30 9 310 17 8 19 3 24 5 58 80

CD34+ Proliferation Assays for Testing GSK3β Inhibitors

Adult hematopoietic stem cells are characterized by the specificexpression of membrane-associated proteins. These surface markers areprovided with an appropriate number appropriate for their molecularweight. This class also includes the molecule which is referred to asCD34 and which serves for the identification, characterization andisolation of adult hematopoietic stem cells. These stem cells canmoreover be isolated from bone marrow, peripheral blood or umbilicalcord blood. These cells have limited viability in in vitro cultures butcan be stimulated to proliferation and differentiation by variousadditions to the culture medium. CD34-positive cells are used here inorder to test the influence of substances on the activity of glycogensynthase kinase 3. For this purpose, in a first step, mononuclear cellsare isolated from umbilical cord blood by differential centrifugationsteps.

For this purpose, umbilical cord blood is diluted 1:4 withphosphate-buffered saline solution. 50 milliliter centrifugation vesselsare charged with 17 milliliters of Ficoll (density 1.077, Ficoll PaquePlus; Pharmacia, catalog number 17-1440-02). 30 milliliters of the 1:4diluted umbilical cord blood are layered thereon and then centrifuged at400×g at room temperature for 30 minutes. The brakes of the centrifugeare disengaged during this. Owing to the centrifugation, the mononuclearcells collect in the interphase. This is removed with the aid of a 30milliliter pipette and transferred into a new 50 millilitercentrifugation vessel, and the volume is then made up to 30 ml withphosphate-buffered saline solution. These cells are centrifuged at 300×gwith the brake engaged at room temperature for 10 minutes. Thesupernatant is discarded and the resulting cell pellet is resuspended in30 milliliters of phosphate-buffered saline solution. These cells areagain centrifuged at 200×g with brake engaged at 20° C. for 15 minutes.

To isolate the CD34-positive cells, the enriched mononuclear cells areresuspended in a concentration of 1×10⁸ cells per 300 microliters ofMACS buffer (0.5% endotoxin-free bovine serum albumin inphosphate-buffered saline solution). 100 microliters of FCR blockingreagent (Miltenyi Biotec, catalog number 130-046-702) and 100microliters of CD34 microbeads (Miltenyi Biotec, catalog number130-046-702) are added. This suspension is incubated at 4° C. for 30minutes. The cells are then diluted with 20 times the volume of MACSbuffer and centrifuged at 300×g for 10 minutes. The supernatant isdiscarded and the cells are resuspended in 500 microliters of MACSbuffer. The cells treated in this way are loaded onto an LS column(Miltenyi Biotec, catalog number 130-042-401) and purified using a MidiMACS magnet (Miltenyi Biotec, catalog number 130-042-303).

The number of CD34-positive cells is determined by counting the cellsusing a Neubauer chamber. The purity of the cells is determined bystandard protocols using the fluorescent activated cell sorting method(Becton Dickinson, BD FACS™ Sample Prep Assistant SPAII Upgrade Kit,catalog number 337642).

To determine the influence of modulating the GSK3 activity,CD34-positive cells are incubated in a 96-well microtiter plate at 37°C. and 5% carbon dioxide for 7 days and then the proliferation rates aredetermined on the basis of the cell counts.

For this purpose, 5000 CD34-positive cells are taken up in 100microliters of IMDM medium (Life Technology, catalog number 12440-046),10% fetal calf serum (Life Technology, catalog number 10082-139) and 20nanograms per milliliter of stem cell factor (R&D, catalog number255-SC-010) in each well of a 96 U-bottom well microtiter plate (GreinerBio-One, catalog number 650 180). In addition, the cells are also mixedwith various concentrations of substances dissolved in dimethylsulfoxide (Sigma Aldrich, catalog number D5879-1L). This entails 4 wellsin each case with the stated cell count of 5000 CD34-positive cells perwell being provided with 10 micromol, 4 wells with 5 micromol, 4 wellswith 2.5 micromol, 4 wells with 1.25 micromol, 4 wells with 0.625micromol, 4 wells with 0.3125 micromol, 4 wells with 0.156 micromol, 4wells with 0.078 micromol and as control 4 wells with 0.1% dimethylsulfoxide as final concentration.

These cells treated in this way are incubated in a cell cultureincubator at 37° C. and 5% carbon dioxide for 7 days. The proliferationrate is determined by renewed counting of the cells using a Neubauercounting chamber, with the cells provided only with the stem cell factorbeing set as 100% value, and all other values being related to thisvalue.

In Vivo Assay

The investigations of the in vivo effect of the compounds according tothe invention take place using 6-week old male C57BL/6 mice (CharlesRiver, Sulzfeld, Germany) weighing 18-22 g. These animals are keptappropriate for the species with 12-hour light and dark cycles underconstant climatic conditions and with water and mouse feed ad libitum.The concentrations of chemotherapeutics used are administered to theanimals in accordance with the manufacturer's statements byintraperitoneal (i.p.) injections in the caudal third of the abdomen.The same procedure is applied to the substances relevant to theinvention. Blood samples are taken from the retrobulbar venous plexususing Pasteur pipettes. The number of neutrophilic granulocytes isdetermined completely automatically using flow cytometry systems.

CYP Inhibition Test

The ability of substances to inhibit CYP1A2, CYP2C8, CYP2C9, CYP2D6 andCYP3A4 in humans is examined using pooled human liver microsomes asenzyme source in the presence of standard substrates (see below) whichform CYP isoform-specific metabolites. The inhibitory effects arestudied at six different concentrations of the test compounds (1.5, 3.1,6.3, 12.5, 25 and 50 μM) and compared to the extent of the CYPisoform-specific metabolite formation of the standard substrates in theabsence of test compounds, and the corresponding IC₅₀ values arecalculated. A standard inhibitor which specifically inhibits a singleCYP isoform serves as control of the results obtained.

Procedure:

The incubation of phenacetin, amodiaquine, diclofenac, dextromethorphanor midazolam with human liver microsomes in the presence of in each casesix different concentrations of a test compound (as potential inhibitor)is carried out on a workstation (Tecan, Genesis, Crailsheim, Germany).Standard incubation mixtures comprise 1.3 mM NADP, 3.3 mM MgCl₂×6 H²O,3.3 mM glucose 6-phosphate, glucose 6-phosphate dehydrogenase (0.4 U/ml)and 100 mM phosphate buffer (pH 7.4) in a total volume of 200 μl. Testcompounds are preferably dissolved in acetonitrile. 96-Well plates areincubated for a defined period of time at 37° C. with pooled human livermicrosomes. The reactions are stopped by addition of 100 μl ofacetonitrile comprising a suitable internal standard. Precipitatedproteins are removed by centrifugation, and the supernatants arecombined and analysed by LC-MS/MS.

Determination of the Solubility Reagents Required:

-   -   PBS buffer pH 6.5: 61.86 g of sodium chloride p.a. (for example        from Merck, Art. No. 1.06404.1000), 39.54 g of sodium dihydrogen        phosphate p.a. (for example from Merck, Art. No. 1.06346.1000)        and 83.35 g of 1 N sodium hydroxide solution (for example from        Bernd Kraft GmbH, Art. No. 01030.4000) are weighed out into a 1        liter measuring flask and made up with water, and the mixture is        stirred for about 1 hour. 500 ml of this solution are        transferred into a 5 liter measuring flask and made up with        water. The pH is adjusted to 6.5 using 1 N sodium hydroxide        solution.    -   Dimethyl sulfoxide (for example from Baker, Art. No. 7157.2500)    -   Distilled water    -   Acetonitrile Chromasolv (for example Riedel-de Haen Art. No.        34851)    -   50% strength formic acid p.a. (for example Fluka Art. No. 09676)

Preparation of the Starting Solution:

At least 1.5 mg of the test substance are weighed out accurately into aWide Mouth 10 mm Screw V-Vial (from Glastechnik Gräfenroda GmbH, Art.No. 8004-WM-H/V 15μ) with fitting screw cap and septum, dimethylsulfoxide is added to give a concentration of 50 mg/ml and the mixtureis vortexed for 30 minutes.

Preparation of the Calibration Solutions:

The required pipetting steps are carried out in a 1.2 ml Deep Well Plate(DWP) with 96 wells (e.g. HJ-Bioanalytik GmbH Art. No. 850289) using aliquid handling robot. The solvent used is a mixture of acetonitrileChromasolv/distilled water 8:2.

Preparation of the Starting Solution for Calibration Solutions (StockSolution):

833 μl of the solvent mixture are added to 10 μl of the initial solution(concentration=600 μg/ml), and the mixture is homogenized. For each testsubstance, 1:100 dilutions are prepared in separate DWPs, and thedilutions for their part are homogenized. One of the 1:100 dilutions isused for preparing the calibration solutions, the second dilution isused for optimizing the MS/MS parameter.

Calibration Solution 5 (600 ng/ml):

270 μl of solvent mixture are added to 30 μl of the stock solution, andthe mixture is homogenized.

Calibration Solution 4 (60 ng/ml):

270 μl of solvent mixture are added to 30 μl of calibration solution 5,and the mixture is homogenized.

Calibration Solution 3 (12 ng/ml):

400 μl of solvent mixture are added to 100 μl of calibration solution 4,and the mixture is homogenized.

Calibration Solution 2 (1.2 ng/ml):

270 μl of solvent mixture are added to 30 μl of calibration solution 3,and the mixture is homogenized.

Calibration Solution 1 (0.6 ng/ml):

150 μl of solvent mixture are added to 150 μl of calibration solution 2,and the mixture is homogenized.

Preparation of the Sample Solutions:

The required pipetting steps are carried out in a 1.2 ml DWP with 96wells (e.g. HJ-Bioanalytik GmbH Art. No. 850289) using a liquid handlingrobot.

1000 μl of PBS buffer pH 6.5 are added to 10.1 μl of the stock solution.

Procedure:

The required pipetting steps are carried out in a 1.2 ml DWP with 96wells (e.g. HJ-Bioanalytik GmbH Art. No. 850289) using a liquid handlingrobot.

Using a temperature-adjustable shaker (e.g. from Eppendorf Thermomixercomfort Art. No. 5355 000.011), the sample solutions prepared in thismanner are shaken at 20° C. and 1400 rpm for 24 hours. From thesesolutions, in each case 180 μl are removed and transferred into Beckmanpolyallomer centrifuge tubes (Art. No. 343621). These solutions arecentrifuged at about 223 000×g for 1 hour (e.g. from Beckman OptimaL-90K Ultracentrifuge with type 42.2 Ti rotor at 42 000 rpm). From eachsample solution, 100 μl of the supernatant are removed and diluted 1:10and 1:1000 with PBS buffer 6.5.

Analysis:

The samples are analysed by HPLC/MS-MS. Quantification is carried outusing a five point calibration curve of the test compound. Thesolubility is expressed in mg/l. Analysis sequence: 1) blank (solventmixture); 2) calibration solution 0.6 ng/ml; 3) calibration solution 1.2ng/ml; 4) calibration solution 12 ng/ml; 5) calibration solution 60ng/ml; 6) calibration solution 600 ng/ml; 7) blank (solvent mixture); 8)sample solution 1:1000; 7) sample solution 1:10.

HPLC/MS-MS Method

HPLC: Agilent 1100, quat. pump (G1311A), autosampler CTC HTS PAL,degasser (G1322A) and column thermostat (G1316A); column: Oasis HLB 20mm×2.1 mm, 25μ; temperature: 40° C.; mobile phase A: water+0.5 ml offormic acid/1; mobile phase B: acetonitrile+0.5 ml of formic acid/1;flow rate: 2.5 ml/min; stop time 1.5 min; gradient: 0 min 95% A, 5% B;ramp: 0-0.5 min 5% A, 95% B; 0.5-0.84 min 5% A, 95% B; ramp: 0.84-0.85min 95% A, 5% B; 0.85-1.5 min 95% A, 5% B.

MS/MS: WATERS Quattro Micro Tandem MS/MS; Z-Spray API interface; HPLC-MSinitial splitter 1:20; measurement in the ESI mode.

For each test substance, the instrument parameters are automaticallyoptimized by injection of the stock solution described further above(second 1:100 dilution) using the MassLynx/QuanOptimize software.

C) EXEMPLARY EMBODIMENTS OF PHARMACEUTICAL COMPOSITIONS

The substances according to the invention can be converted intopharmaceutical preparations in the following ways:

Tablet: Composition:

100 mg of the compound of Example 1, 50 mg of lactose (monohydrate), 50mg of maize starch, 10 mg of polyvinylpyrrolidone (PVP 25) (from BASF,Germany) and 2 mg of magnesium stearate.

Tablet weight 212 mg. Diameter 8 mm, radius of curvature 12 mm.

Production:

The mixture of the compound of Example 1, lactose and starch isgranulated with a 5% strength solution (m/m) of the PVP in water. Thegranules are dried and then mixed with magnesium stearate for 5 min.This mixture is compressed with a conventional tablet press (see abovefor format of the tablet).

Oral Suspension: Composition:

1000 mg of the compound of Example 1, 1000 mg of ethanol (96%), 400 mgof Rhodigel (xanthan gum) (from FMC, USA) and 99 g of water.

10 ml of oral suspension correspond to a single dose of 100 mg of thecompound according to the invention.

Production:

The Rhodigel is suspended in ethanol, and the compound of Example 1 isadded to the suspension. The water is added while stirring. The mixtureis stirred for about 6 h until the swelling of the Rhodigel is complete.

Solution which can be Administered Intravenously:

Composition:

1 mg of the compound of Example 1, 15 g of polyethylene glycol 400 and250 g of water for injections.

Production:

The compound of Example 1 is dissolved together with polyethylene glycol400 in the water by stirring. This solution is sterilized by filtration(pore diameter 0.22 μm) and dispensed under aseptic conditions intoheat-sterilized infusion bottles. These are closed with infusionstoppers and crimped caps.

1. A compound of the formula

in which either U represents N, V represents CR¹², W represents CH, Arepresents CR¹⁵, or U represents CH, V represents CR¹², W represents N,A represents CR¹⁵, or U represents CR¹⁶, V represents N, W representsCR^(L7), A represents N, where R¹² represents hydrogen, hydroxyl, amino,hydroxycarbonyl, aminocarbonyl, trifluoromethyl, trifluoromethoxy,cyano, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkylcarbonyl,C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,C₁-C₄-alkylcarbonylamino, C₁-C₄-alkylsulfonyl-amino, 5- or 6-memberedheterocyclylcarbonyl, —CH₂R¹³ or —CH₂CH₂R¹⁴, where heterocyclylcarbonylmay be substituted by 1 to 3 substituents, where the substituentsindependently of one another are selected from the group consisting ofhalogen, oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl,and where alkoxy, alkylamino, alkylcarbonyl, alkoxycarbonyl,alkylaminocarbonyl, alkylcarbonylamino and alkylsulfonylamino may besubstituted by a substituent, where the substituent is selected from thegroup consisting of hydroxyl, amino, hydroxycarbonyl, aminocarbonyl,C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino, 5- or 6-memberedheterocyclyl and phenyl, where phenyl may be substituted by 1 to 3substituents, where the substituents independently of one another areselected from the group consisting of halogen, cyano, trifluoromethyl,trifluoromethoxy, aminocarbonyl, C₁-C₄-alkyl, C₁-C₄-alkoxy,C₁-C₄-alkylamino, C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino, and whereheterocyclyl may be substituted by 1 to 3 substituents, where thesubstituents independently of one another are selected from the groupconsisting of halogen, oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl,and where R¹³ represents hydroxyl, amino, cyano, hydroxycarbonyl,aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino,C₃-C₆-cycloalkylamino or 5- or 6-membered heterocyclyl, where alkoxy,alkylamino, alkoxycarbonyl, alkylaminocarbonyl and alkylcarbonylaminomay be substituted by a substituent, where the substituent is selectedfrom the group consisting of hydroxyl, amino, hydroxycarbonyl,aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino, and whereheterocyclyl may be substituted by 1 to 3 substituents, where thesubstituents independently of one another are selected from the groupconsisting of halogen, oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl,and where R¹⁴ represents hydroxyl, amino, cyano, hydroxycarbonyl,aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino or 5- or 6-memberedheterocyclyl, where alkoxy, alkylamino, alkoxycarbonyl,alkylaminocarbonyl and alkylcarbonylamino may be substituted by asubstituent, where the substituent is selected from the group consistingof hydroxyl, amino, hydroxycarbonyl, aminocarbonyl, C₁-C₄-alkoxy,C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl andC₁-C₄-alkylcarbonylamino, and where heterocyclyl may be substituted by 1to 3 substituents, where the substituents independently of one anotherare selected from the group consisting of halogen, oxo, C₁-C₄-alkyl,C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkylcarbonyl,C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl, R¹⁵ representshydrogen, halogen, cyano, trifluoromethyl, C₁-C₃-alkyl, methoxy,methylthio or cyclopropyl, R¹⁶ represents hydrogen or methyl, R¹⁷represents hydrogen or methyl, R¹ represents a group of the formula

where is the point of attachment to the heterocycle, and R³ represents2-pyridyl, pyrimid-2-yl, 2-aminopyrimid-4-yl,2-(mono-C₁-C₄-alkylamino)pyrimid-4-yl,2-(mono-C₃-C₄-cycloalkylamino)pyrimid-4-yl, pyridazin-3(2H)-on-6-yl,1,3-oxazol-2-yl, 1,3-oxazol-4-yl, 1,2,4-oxadiazol-3-yl,1,2,3-oxadiazol-4-yl, 1,3-thiazol-2-yl, 1,3-thiazol-4-yl,1H-1,2,4-triazol-5-yl, 2,4-dihydro-3H-1,2,4-triazol-3-on-5-yl or1,2-pyrazol-5-yl, where 2-pyridyl, pyrimid-2-yl, 1,3-oxazol-2-yl,1,3-oxazol-4-yl, 1,3-thiazol-2-yl and 1,3-thiazol-4-yl are substitutedby 1 or 2 substituents, where the substituents independently of oneanother are selected from the group consisting of halogen, cyano, nitro,amino, trifluoromethyl, trifluoromethoxy, aminocarbonyl,trifluoromethylcarbonyl, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,C₃-C₄-cycloalkylamino, C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl and C₃-C₆-cycloalkylcarbonyl, where alkyl,alkoxy, alkylamino, alkylcarbonyl, alkoxycarbonyl, alkylaminocarbonyland cycloalkylcarbonyl may be substituted by a substituent, where thesubstituent is selected from the group consisting of halogen, cyano,hydroxyl, amino, trifluoromethyl and C₃-C₆-cycloalkyl, and where2-aminopyrimid-4-yl, 2-(mono-C₁-C₄-alkylamino)pyrimid-4-yl,2-(mono-C₃-C₄-cycloalkylamino)pyrimid-4-yl, pyridazin-3(2H)-on-6-yl,1,2,4-oxadiazol-3-yl, 1,2,3-oxadiazol-4-yl, 1H-1,2,4-triazol-5-yl,2,4-dihydro-3H-1,2,4-triazol-3-on-5-yl and 1,2-pyrazol-5-yl may besubstituted by a substituent, where the substituent is selected from thegroup consisting of halogen, cyano, nitro, amino, trifluoromethyl,trifluoromethoxy, aminocarbonyl, trifluoromethylcarbonyl, C₁-C₄-alkyl,C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₃-C₄-cycloalkylamino,C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl andC₃-C₆-cycloalkylcarbonyl, R² represents C₆-C₁₀-aryl or 5- to 10-memberedheteroaryl, where aryl and heteroaryl may be substituted by 1 to 3substituents, where the substituents independently of one another areselected from the group consisting of hydroxyl, hydroxymethyl, amino,halogen, cyano, trifluoromethyl, trifluoromethoxy, aminocarbonyl,C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkoxymethyl, C₁-C₄-alkylamino,C₁-C₄-alkylaminomethyl, C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino, C₁-C₄-alkylsulfonyl,C₁-C₄-alkylsulfonylamino, C₁-C₄-alkylaminosulfonyl, phenyl, benzyloxy,5- or 6-membered heterocyclyl, 5- or 6-membered heterocyclylcarbonyl, 5-or 6-membered heterocyclylmethyl and 5- or 6-membered heteroaryl, wherephenyl, benzyloxy, heterocyclyl, heterocyclylcarbonyl,heterocyclylmethyl and heteroaryl may be substituted by 1 to 3substituents, where the substituents independently of one another areselected from the group consisting of halogen, cyano, trifluoromethyl,trifluoromethoxy, aminocarbonyl, C₁-C₄-alkyl, C₁-C₄-alkoxy,C₁-C₄-alkylamino, C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino, or two of thesubstituents on aryl together with the carbon atoms to which they areattached form a 1,3-dioxolane or 1,4-dioxane, or one of its salts. 2.The compound as claimed in claim 1, characterized in that either Urepresents N, V represents CR¹², W represents CH, A represents CR¹⁵, orU represents CH, V represents CR¹², W represents N, A represents CR¹⁵,or U represents CR¹⁶, V represents N, W represents CR¹⁷, A represents N,where R¹² represents hydrogen, hydroxycarbonyl, aminocarbonyl,C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkylcarbonyl,C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,C₁-C₄-alkylcarbonylamino, 5- or 6-membered heterocyclylcarbonyl, —CH₂R¹³or —CH₂CH₂R¹⁴, where heterocyclylcarbonyl may be substituted by 1 to 3substituents, where the substituents independently of one another areselected from the group consisting of oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy,C₁-C₄-alkylamino, C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl andC₁-C₄-alkylaminocarbonyl, and where alkoxy, alkylamino, alkylcarbonyl,alkoxycarbonyl, alkylaminocarbonyl and alkylcarbonylamino may besubstituted by a substituent, where the substituent is selected from thegroup consisting of hydroxyl, amino, hydroxycarbonyl, aminocarbonyl,C₁-C₄-alkoxy, C₁-C₄-alkylamino and 5- or 6-membered heterocyclyl, whereheterocyclyl may be substituted by 1 to 2 substituents, where thesubstituents independently of one another are selected from the groupconsisting of oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl,and where R¹³ represents hydroxyl, amino, hydroxycarbonyl,aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino,C₃-C₆-cycloalkylamino or 5- or 6-membered heterocyclyl, where alkoxy,alkylamino, alkoxycarbonyl, alkylaminocarbonyl and alkylcarbonylaminomay be substituted by a substituent, where the substituent is selectedfrom the group consisting of hydroxyl, amino, hydroxycarbonyl,aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl and C₁-C₄-alkylcarbonylamino, and whereheterocyclyl may be substituted by 1 to 2 substituents, where thesubstituents independently of one another are selected from the groupconsisting of oxo, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl,and where R¹⁴ represents hydroxyl, amino, hydroxycarbonyl,aminocarbonyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl,C₁-C₄-alkylaminocarbonyl, C₁-C₄-alkylcarbonylamino or 5- or 6-memberedheterocyclyl, where alkoxy, alkylamino, alkoxycarbonyl,alkylaminocarbonyl and alkylcarbonylamino may be substituted by asubstituent, where the substituent is selected from the group consistingof hydroxyl, amino, hydroxycarbonyl, aminocarbonyl, C₁-C₄-alkoxy,C₁-C₄-alkylamino, C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl andC₁-C₄-alkylcarbonylamino, and where heterocyclyl may be substituted by 1to 2 substituents, where the substituents independently of one anotherare selected from the group consisting of oxo, C₁-C₄-alkyl,C₁-C₄-alkoxy, C₁-C₄-alkylamino, C₁-C₄-alkylcarbonyl,C₁-C₄-alkoxycarbonyl and C₁-C₄-alkylaminocarbonyl, R¹⁵ representshydrogen, halogen, cyano or trifluoromethyl, R¹⁶ represents hydrogen ormethyl, R¹⁷ represents hydrogen or methyl, R¹ represents a group of theformula

where is the point of attachment to the heterocycle, and R³ represents2-pyridyl, pyrimid-2-yl, 2-aminopyrimid-4-yl, 1,3-oxazol-2-yl,1,3-oxazol-4-yl, 1,2,4-oxadiazol-3-yl, 1,2,3-oxadiazol-4-yl,1,3-thiazol-2-yl or 1,3-thiazol-4-yl, where 2-pyridyl, pyrimid-2-yl,1,3-oxazol-2-yl, 1,3-oxazol-4-yl, 1,3-thiazol-2-yl and 1,3-thiazol-4-ylare substituted by 1 or 2 substituents, where the substituentsindependently of one another are selected from the group consisting ofhalogen, cyano, nitro, amino, trifluoromethyl, trifluoromethoxy,aminocarbonyl, trifluoromethylcarbonyl methyl, ethyl, methoxy, ethoxy,C₁-C₄-alkylamino, methylcarbonyl, ethylcarbonyl, cyclopropylcarbonyl,methoxycarbonyl and ethoxycarbonyl, and where 2-aminopyrimid-4-yl,1,2,4-oxadiazol-3-yl and 1,2,3-oxadiazol-4-yl may be substituted by asubstituent, where the substituent is selected from the group consistingof halogen, cyano, nitro, amino, trifluoromethyl, trifluoromethoxy,aminocarbonyl, trifluoromethylcarbonyl, methyl, ethyl, methoxy, ethoxy,C₁-C₄-alkylamino, methylcarbonyl, ethylcarbonyl, cyclopropylcarbonyl,methoxycarbonyl and ethoxycarbonyl, R² represents C₆-C₁₀-aryl, thienyl,furyl, pyrrolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,imidazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, indolyl,indazolyl, quinolinyl, benzofuranyl or benzoxazolyl, where aryl,thienyl, furyl, pyrrolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,imidazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, indolyl,indazolyl, quinolinyl, benzofuranyl and benzoxazolyl may be substitutedby 1 to 3 substituents, where the substituents independently of oneanother are selected from the group consisting of hydroxyl,hydroxymethyl, amino, halogen, cyano, trifluoromethyl, trifluoromethoxy,aminocarbonyl, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkoxymethyl,C₁-C₄-alkylamino, C₁-C₄-alkylaminomethyl, C₁-C₄-alkylcarbonyl,C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,C₁-C₄-alkylcarbonylamino, C₁-C₄-alkylsulfonyl, C₁-C₄-alkylsulfonylamino,C₁-C₄-alkylaminosulfonyl, phenyl, benzyloxy, 5- or 6-memberedheterocyclyl, 5- or 6-membered heterocyclylcarbonyl, 5- or 6-memberedheterocyclylmethyl and 5- or 6-membered heteroaryl, where phenyl,benzyloxy, heterocyclyl, heterocyclylcarbonyl, heterocyclylmethyl andheteroaryl may be substituted by 1 to 3 substituents, where thesubstituents independently of one another are selected from the groupconsisting of halogen, cyano, trifluoromethyl, trifluoromethoxy,aminocarbonyl, C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-alkylamino,C₁-C₄-alkylcarbonyl, C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl andC₁-C₄-alkylcarbonylamino, or one of its salts.
 3. The compound asclaimed in claim 1, characterized in that R² is phenyl optionallysubstituted by 1 to 2 substituents, where the substituents independentlyof one another are selected from the group consisting of halogen,trifluoromethyl, trifluoromethoxy, aminocarbonyl, C₁-C₄-alkyl,C₁-C₄-alkoxy, C₁-C₄-alkoxycarbonyl, C₁-C₄-alkylaminocarbonyl,pyrrolidinyl, piperidinyl, morpholinyl and morpholinylcarbonyl.
 4. Aprocess for preparing a compound of the formula (I) or one of its saltsas claimed in claim 1, characterized in that [A] a compound of theformula

in which A, U, V, W and R² have the meaning given in claim 1, and X¹represents halogen, preferably chlorine or fluorine, is reacted withcompounds of the formulaR¹—H  (III), in which R¹ has the meaning given in claim 1, or [B] acompound of the formula

in which R¹ has the meaning given in claim 1, and A represents CR¹⁵,where R¹⁵ has the meaning given in claim 1, U represents N, V representsCR¹², where R¹² has the meaning given in claim 1, W represents CH, X²represents iodine, bromine, chlorine or trifluoromethanesulfonyl,preferably iodine or bromine, is reacted under Suzuki couplingconditions with a compound of the formulaQ—R²  (V), in which R² has the meaning given in claim 1, and Qrepresents —B(OH)₂, a boronic acid ester, preferably boronic acidpinacolate, or —BF₃ ⁻K⁺, to give a compound of the formula

in which R¹ and R² have the meaning given in claim 1, and A representsCR¹⁵, where R¹⁵ has the meaning given in claim 1, U represents N, Vrepresents CR¹², where R¹² has the meaning given in claim 1, Wrepresents CH.
 5. A pharmaceutical composition, comprising a compound asclaimed in claim 1 in combination with an inert non-toxicpharmaceutically acceptable auxiliary.
 6. The pharmaceutical compositionas claimed in claim 5 for the treatment and/or prophylaxis ofhematological disorders.
 7. A method for treating a hematologic disordercomprising administering to a human or animal in need thereof atherapeutically effective amount of a compound as claimed in claim
 1. 8.A method for the ex vivo expansion of adult hematopoietic stem cellsfrom bone marrow, from peripheral blood or umbilical cord blood,characterized in that an effective amount of a compound as claimed inclaim 1 is added.